Advantages and disadvantages of detection methods.
\\n\\n
Dr. Pletser’s experience includes 30 years of working with the European Space Agency as a Senior Physicist/Engineer and coordinating their parabolic flight campaigns, and he is the Guinness World Record holder for the most number of aircraft flown (12) in parabolas, personally logging more than 7,300 parabolas.
\\n\\nSeeing the 5,000th book published makes us at the same time proud, happy, humble, and grateful. This is a great opportunity to stop and celebrate what we have done so far, but is also an opportunity to engage even more, grow, and succeed. It wouldn't be possible to get here without the synergy of team members’ hard work and authors and editors who devote time and their expertise into Open Access book publishing with us.
\\n\\nOver these years, we have gone from pioneering the scientific Open Access book publishing field to being the world’s largest Open Access book publisher. Nonetheless, our vision has remained the same: to meet the challenges of making relevant knowledge available to the worldwide community under the Open Access model.
\\n\\nWe are excited about the present, and we look forward to sharing many more successes in the future.
\\n\\nThank you all for being part of the journey. 5,000 times thank you!
\\n\\nNow with 5,000 titles available Open Access, which one will you read next?
\\n\\nRead, share and download for free: https://www.intechopen.com/books
\\n\\n\\n\\n
\\n"}]',published:!0,mainMedia:null},components:[{type:"htmlEditorComponent",content:'
Preparation of Space Experiments edited by international leading expert Dr. Vladimir Pletser, Director of Space Training Operations at Blue Abyss is the 5,000th Open Access book published by IntechOpen and our milestone publication!
\n\n"This book presents some of the current trends in space microgravity research. The eleven chapters introduce various facets of space research in physical sciences, human physiology and technology developed using the microgravity environment not only to improve our fundamental understanding in these domains but also to adapt this new knowledge for application on earth." says the editor. Listen what else Dr. Pletser has to say...
\n\n\n\nDr. Pletser’s experience includes 30 years of working with the European Space Agency as a Senior Physicist/Engineer and coordinating their parabolic flight campaigns, and he is the Guinness World Record holder for the most number of aircraft flown (12) in parabolas, personally logging more than 7,300 parabolas.
\n\nSeeing the 5,000th book published makes us at the same time proud, happy, humble, and grateful. This is a great opportunity to stop and celebrate what we have done so far, but is also an opportunity to engage even more, grow, and succeed. It wouldn't be possible to get here without the synergy of team members’ hard work and authors and editors who devote time and their expertise into Open Access book publishing with us.
\n\nOver these years, we have gone from pioneering the scientific Open Access book publishing field to being the world’s largest Open Access book publisher. Nonetheless, our vision has remained the same: to meet the challenges of making relevant knowledge available to the worldwide community under the Open Access model.
\n\nWe are excited about the present, and we look forward to sharing many more successes in the future.
\n\nThank you all for being part of the journey. 5,000 times thank you!
\n\nNow with 5,000 titles available Open Access, which one will you read next?
\n\nRead, share and download for free: https://www.intechopen.com/books
\n\n\n\n
\n'}],latestNews:[{slug:"stanford-university-identifies-top-2-scientists-over-1-000-are-intechopen-authors-and-editors-20210122",title:"Stanford University Identifies Top 2% Scientists, Over 1,000 are IntechOpen Authors and Editors"},{slug:"intechopen-authors-included-in-the-highly-cited-researchers-list-for-2020-20210121",title:"IntechOpen Authors Included in the Highly Cited Researchers List for 2020"},{slug:"intechopen-maintains-position-as-the-world-s-largest-oa-book-publisher-20201218",title:"IntechOpen Maintains Position as the World’s Largest OA Book Publisher"},{slug:"all-intechopen-books-available-on-perlego-20201215",title:"All IntechOpen Books Available on Perlego"},{slug:"oiv-awards-recognizes-intechopen-s-editors-20201127",title:"OIV Awards Recognizes IntechOpen's Editors"},{slug:"intechopen-joins-crossref-s-initiative-for-open-abstracts-i4oa-to-boost-the-discovery-of-research-20201005",title:"IntechOpen joins Crossref's Initiative for Open Abstracts (I4OA) to Boost the Discovery of Research"},{slug:"intechopen-hits-milestone-5-000-open-access-books-published-20200908",title:"IntechOpen hits milestone: 5,000 Open Access books published!"},{slug:"intechopen-books-hosted-on-the-mathworks-book-program-20200819",title:"IntechOpen Books Hosted on the MathWorks Book Program"}]},book:{item:{type:"book",id:"7613",leadTitle:null,fullTitle:"Research Trends and Challenges in Smart Grids",title:"Research Trends and Challenges in Smart Grids",subtitle:null,reviewType:"peer-reviewed",abstract:"This book introduces the most promising enabling technologies and methodologies for smart grids. It not only focuses on technological breakthroughs and roadmaps in implementing these technologies, but also presents the much-needed sharing of best practices, demonstrating the potential role of smart grid functions in improving the technical, economic, and environmental performance of modern power distribution systems. 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From March 2002 to\nOctober 2014 he has been Assistant Professor of Electric Power\nSystems at the Department of Engineering, Faculty of Engineering\nof the University of Sannio. From February 2011 to December\n2013 he was the Scientific Director of the Bureau of the Research Centre on Pure and\nApplied Mathematics at the Department of Engineering, University of Sannio. On\nOctober 2014 he obtained the National Scientific Qualification of Full Professor in\nElectrical Energy Engineering. Since November 2014 he has been Associate Professor\nof Electric Power Systems at the Department of Engineering of University of Sannio.\nHe is the Editor in Chief of Technology and Economics of Smart Grids and Sustainable\nEnergy, Springer Nature, Chair of the PES-IEEE Task Force on Enabling Paradigms\nfor High-Performance Computing in Wide Area Monitoring Protective and Control\nSystems, and Chair of the IEEE PES Awards and Recognition Committee.",institutionString:"University of Sannio, Benevento",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"1",totalChapterViews:"0",totalEditedBooks:"1",institution:{name:"University of Sannio",institutionURL:null,country:{name:"Italy"}}}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,coeditorOne:{id:"39249",title:"Dr.",name:"Ahmed F.",middleName:null,surname:"Zobaa",slug:"ahmed-f.-zobaa",fullName:"Ahmed F. 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\r\n\tThe use of dyes and pigments is narrowly associated with everyday life. Since ancient times, people have been using various types of dyes and pigments for both aesthetic and practical applications. Typically, the coloration of various materials e.g. textiles, clay, plastics, etc. has been their main purpose. Yet, the scope of contemporary dyes and pigments has become significantly broader and there is constant interest in new products fulfilling numerous requirements parallel to their ability to act as colorants. This trend has led to the development of functional dyes.
\r\n\r\n\tIn recent years, novel dyes and pigments with hi-tech applications have been developed and there is a continuous demand for new products with better properties and/or broader application scope. Of particular interest is the development of dyes and pigments with environment-responsive aptitudes i.e. products that can undergo some structural modification as a result of external stimuli e.g. light, heat, pressure, pH-changes, etc. These stimuli-responsive functional dyes have in turn found application in sensor technologies, optical data storage, molecular switches, etc. Acknowledging these facts, this book aims to cover current state-of-the-art research and development in the remarkably important area of environment-responsive (multi)functional dyes and pigments.
",isbn:"978-1-83968-615-3",printIsbn:"978-1-83968-614-6",pdfIsbn:"978-1-83968-616-0",doi:null,price:0,priceEur:0,priceUsd:0,slug:null,numberOfPages:0,isOpenForSubmission:!1,hash:"624f533946a159bc8a03f109c2e1dc91",bookSignature:"Dr. Raffaello Papadakis",publishedDate:null,coverURL:"https://cdn.intechopen.com/books/images_new/10203.jpg",keywords:"Fluorescent Dyes, PH-Sensitive Dyes, Solvatochromism, Solvent Polarity Indicators, Chromic Betaines, Viscosity, Charge-Transfer Complexes, Spectroscopy, Piezochromism, Optoelectronics, Photochromism, Molecular Switches",numberOfDownloads:62,numberOfWosCitations:0,numberOfCrossrefCitations:0,numberOfDimensionsCitations:0,numberOfTotalCitations:0,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"August 28th 2020",dateEndSecondStepPublish:"September 25th 2020",dateEndThirdStepPublish:"November 24th 2020",dateEndFourthStepPublish:"February 12th 2021",dateEndFifthStepPublish:"April 13th 2021",remainingDaysToSecondStep:"5 months",secondStepPassed:!0,currentStepOfPublishingProcess:5,editedByType:null,kuFlag:!1,biosketch:"Chemical Engineer with working experience at the Institute of Molecular Sciences, CNRS/Aix-Marseille University in the research group of Dr. Thierry Tron, and at Uppsala University in the research group of Dr. Henrik Ottosson. Currently a Senior Research Scientist at Tdb Labs, Uppsala, Sweden.",coeditorOneBiosketch:null,coeditorTwoBiosketch:null,coeditorThreeBiosketch:null,coeditorFourBiosketch:null,coeditorFiveBiosketch:null,editors:[{id:"251885",title:"Dr.",name:"Raffaello",middleName:null,surname:"Papadakis",slug:"raffaello-papadakis",fullName:"Raffaello Papadakis",profilePictureURL:"https://mts.intechopen.com/storage/users/251885/images/system/251885.jpg",biography:"Raffaello Papadakis is a Chemical Engineer (MEng 2005) majoring in organic chemical technology and polymer science and technology. He started his PhD in the field of physical organic chemistry in 2006 under the supervision of Prof. (Emer.) Dr. Athanase Tsolomitis (National Technical University of Athens, Greece) and graduated in 2010. During his PhD he concentrated on the synthesis of solvatochromic probes and molecular switches. He later on spent two years in Marseille, France (September 2010–January 2013) working as a postodoctoral researcher at the Institute of Molecular Sciences, CNRS/Aix-Marseille University, in the field of water oxidation catalysts in the research group of Dr. Thierry Tron before moving to to Uppsala, Sweden in 2014. There he joined the group of Dr. Henrik Ottosson (Uppsala University) and he worked as a postdoc researcher and later as a researcher (Forskare) focusing on excited state (anti)aromaticity and graphene photochemistry-related research. His current research interests revolve around physical organic and materials chemistry with an emphasis on the chemistry and photochemistry of graphene and novel covalent organic frameworks as well as polymer chemistry. He is the author and coauthor of 24 scientific research papers, two book chapters and he has more than 35 contributions in international conference proceedings. Furthermore, he is an active referee of scientific peer-reviewed papers of world-class chemistry journals and he has acted as a scientific expert evaluating international research-grant proposals. Currently he works as a Senior Research Scientist at TdB Labs AB (Sweden) and specializes in polysaccharide modifications and derivatization placing particular interest in fluorescent dye polysaccharide-functionalizations.",institutionString:"TdB Labs",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"2",totalChapterViews:"0",totalEditedBooks:"0",institution:null}],coeditorOne:null,coeditorTwo:null,coeditorThree:null,coeditorFour:null,coeditorFive:null,topics:[{id:"8",title:"Chemistry",slug:"chemistry"}],chapters:[{id:"74730",title:"Treatment of Textile Dyeing Waste Water Using TiO2/Zn Electrode by Spray Pyrolysis in Electrocoagulation Process",slug:"treatment-of-textile-dyeing-waste-water-using-tio2-zn-electrode-by-spray-pyrolysis-in-electrocoagula",totalDownloads:63,totalCrossrefCites:0,authors:[null]}],productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"},personalPublishingAssistant:{id:"247865",firstName:"Jasna",lastName:"Bozic",middleName:null,title:"Ms.",imageUrl:"https://mts.intechopen.com/storage/users/247865/images/7225_n.jpg",email:"jasna.b@intechopen.com",biography:"As an Author Service Manager, my responsibilities include monitoring and facilitating all publishing activities for authors and editors. 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Venkateswarlu",coverURL:"https://cdn.intechopen.com/books/images_new/371.jpg",editedByType:"Edited by",editors:[{id:"58592",title:"Dr.",name:"Arun",surname:"Shanker",slug:"arun-shanker",fullName:"Arun Shanker"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"878",title:"Phytochemicals",subtitle:"A Global Perspective of Their Role in Nutrition and Health",isOpenForSubmission:!1,hash:"ec77671f63975ef2d16192897deb6835",slug:"phytochemicals-a-global-perspective-of-their-role-in-nutrition-and-health",bookSignature:"Venketeshwer Rao",coverURL:"https://cdn.intechopen.com/books/images_new/878.jpg",editedByType:"Edited by",editors:[{id:"82663",title:"Dr.",name:"Venketeshwer",surname:"Rao",slug:"venketeshwer-rao",fullName:"Venketeshwer Rao"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"4816",title:"Face Recognition",subtitle:null,isOpenForSubmission:!1,hash:"146063b5359146b7718ea86bad47c8eb",slug:"face_recognition",bookSignature:"Kresimir Delac and Mislav Grgic",coverURL:"https://cdn.intechopen.com/books/images_new/4816.jpg",editedByType:"Edited by",editors:[{id:"528",title:"Dr.",name:"Kresimir",surname:"Delac",slug:"kresimir-delac",fullName:"Kresimir Delac"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"3621",title:"Silver Nanoparticles",subtitle:null,isOpenForSubmission:!1,hash:null,slug:"silver-nanoparticles",bookSignature:"David Pozo Perez",coverURL:"https://cdn.intechopen.com/books/images_new/3621.jpg",editedByType:"Edited by",editors:[{id:"6667",title:"Dr.",name:"David",surname:"Pozo",slug:"david-pozo",fullName:"David Pozo"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}}]},chapter:{item:{type:"chapter",id:"58995",title:"Current and Emerging Technologies for Rapid Detection of Pathogens",doi:"10.5772/intechopen.73178",slug:"current-and-emerging-technologies-for-rapid-detection-of-pathogens",body:'\nFoodborne pathogens, which are widely responsible for many foodborne diseases, constitute a serious threat to human health. In recent years, foodborne and waterborne pathogenic microorganisms have caused numerous epidemic diseases in the world [1]. Salmonella, Shiga bacillus, Escherichia coli O157:H7, Bacillus cereus, Staphylococcus aureus, and Listeria monocytogenes are the primary pathogens that are responsible for most foodborne disease [2, 3, 4, 5, 6]. Centers for Disease Control and Prevention (CDC) reported that approximately 73,000 cases of foodborne disease occur annually. In 2013, a total of 19,056 are infected with foodborne pathogen, of which 4200 are hospitalizations, and 80 are deaths in the United States [7]. The foodborne diseases are even high prevalence in many developing countries. Worldwide, there are 600 million foodborne illnesses with 420,000 deaths in 2010, which is estimated by the World Health Organization (WHO) Foodborne Disease Burden Epidemiology Reference Group (FERG). A great proportion of these cases are due to the contamination of raw or undercooked foods and drinking water [4, 6, 8, 9]. Hence, it is urgent to detect foodborne pathogens in order to control foodborne pathogen spread and reduce foodborne disease occurrence.
\nCurrently, culture-based bacterial isolation and identification are the “gold standard” methods for laboratory detection of foodborne pathogens [10]. However, they suffer from time consumption, which requires 2–3 days for initial culture and enrichment, and more than 1 week for confirming the target pathogenic bacteria [11, 12]. Moreover, it requires expensive instruments and professional technicians and remains problematic due to the lack of phenotypic characteristics to distinguish between generic pathogens, which may largely restrict its application. It is evident that culture and colony-counting methods are inadequate for rapid detection of foodborne pathogens, especially for reduce foodborne disease occurrence. The frequent outbreak of foodborne diseases and the economic and social implications indicate that analytical methodologies that can rapidly detect and identify pathogens are urgently needed. As such, many researchers devote themselves to developing more advanced detection methods that can identify pathogens accurately and rapidly in a timely manner in the food industry [13, 14, 15, 16, 17, 18, 19, 20].
\nIn this chapter, we summarize the recent trends, developments, advantages, and disadvantages (listed in Table 1) about rapid detection of pathogens based on nucleic acid, antibodies, and aptamers and then give a perspective on the future directions of rapid analysis of pathogens.
\nMethod | \nAdvantages | \nDisadvantages | \nSensitivity | \nRef. | \n
---|---|---|---|---|
Real-time PCRa | \n—Amplification can be monitored at real time —Confirmation of specific amplification by melting curve —Accurate quantification | \n—Difficulty in multiplex assay —Need skilled person and support —False-positive results | \n10 CFU/mL | \n[25] | \n
Multiplex PCRa | \n—Highly efficient (detection of several pathogens at a time) —Systematic (suitable for detection of groups of pathogens) | \n—Difficulty in distinguishing live and dead cells —Requires post-PCR processing of products (electrophoresis) —Need skilled person and support —Costs more than culture-based methods and ELISA | \n1 CFU/mL | \n[26] | \n
Antibody-based method (ELISAb and LFIAc) | \n—More rapid than culture-based methods (1~2 h vs. 5~7 d) —Can be automated to reduce assay time and manual labor input —Able to handle large numbers of samples —Convenient and suitable for the on-site testing | \n—Difficulty to differentiate damaged or stressed cells —Need for pre-enrichment —High cross-reactivity with close antigens in bacteria | \n60 CFU/mL | \n[35] | \n
Aptamer-based method (optical and electrochemical methods) | \n—Inexpensive, stable, and can be chemically synthesized than antibody —Time-saving (2 h vs. 5~7 d of culture-based methods) —Automated to reduce manual labor input —High throughput —Multiplex assays | \n—High false positive —Difficulty in detecting damaged or stressed cells —Need for pre-enrichment —Possibility of cross contamination | \n1.5 CFU/mL | \n[78] | \n
Advantages and disadvantages of detection methods.
a PCR, polymerase chain reaction; b ELISA, enzyme-linked immunosorbent assay; c LFIA, lateral flow immunoassays.
Culture- and colony-based methods are the standard methods for the detection of pathogens. They rely on the ability of microorganisms to multiply to visible colonies [21]. The major drawbacks of these microbiological methods are their labor intensiveness and time consumption as it usually takes 2–3 days for initial results and up to 7–10 days for confirmation. In comparison, nucleic acid-based assays can greatly shorten the testing time.
\nReal-time PCR technology is a reliable method in identification and quantitative detection of bacteria due to its accuracy, rapidity, specificity, and low detection limit. In addition, it is a promising alternative approach to estimating the number of bacteria [22, 23]. For example, Gyawali et al. [22] presented a specific and sensitive real-time PCR method to detect Ancylostoma caninum ova in wastewater matrices. This method exhibited high sensitivity with the ability to detect Ancylostoma caninum DNA up to dilution of 10−4 (equivalent to 500 fg) consistently. Moreover, the precise copy number of a specific nucleic acid sequence can be quantified with the real-time PCR technique based on a calibration curve created with known concentrations of DNA [24, 25]. Gokduman et al. [25] established a recombinant plasmid-based quantitative real-time PCR assay for Salmonella enterica serotypes with the detection limit of 10 CFU/mL. Obviously, the real-time quantitative PCR has already been a promising quantitative method for the quantitative detection of bacteria, due to its lower cost than that of culture-based method.
\nMultiplex PCR, also known as multiple primer PCR, which is a PCR reaction system with two or more primers, can amplify a plurality of nucleic acid fragments in a system. Compared to other methods, multiplex PCR is very useful as it allows the simultaneous detection of several pathogenic bacteria by introducing different primers to amplify DNA regions coding for specific genes of each bacterial strain targeted [26]. Methods for multiplexing PCR have considerably improved over the last years, thereby decreasing genotyping costs and increasing throughput. Examples of multiplex PCR technique for the simultaneous detection pathogens include multiplex PCR assay for rapid and simultaneous detection of Escherichia coli O157:H7, Salmonella, Staphylococcus aureus, Listeria monocytogenes, and Vibrio parahaemolyticus [27]; simultaneous detection of bacteria of the genus Listeria, Listeria monocytogenes, and major serotypes and epidemic clones of Listeria monocytogenes [28]; and simultaneous detection of Escherichia coli O157:H7 and Listeria monocytogenes [29]. Multiplex real-time PCR-based assay can rapidly detect 25 clinically important pathogens directly from whole blood in <6 h [30].
\nAntibodies are a unique natural family of immune system-related glycoproteins known as immunoglobulins, produced by differentiated B cells in response to the attendant of an immunogen during an immune response. Because of the specific interactions and the extremely high equilibrium association constants (1010/M and greater) attainable between an antibody and its corresponding antigen, antibodies are employed as an excellent biorecognition element for the highly sensitive and selective immunoassays [31]. Their utilization in biosensors brings new tools for analysis in the biochemical, clinical, and environmental fields. Without exception, antibody-based assays such as enzyme-linked immunosorbent assay (ELISA), lateral flow immunoassay (LFIA), and so on are very popular for the detection of pathogens.
\nELISA-based approaches are the most prevalent antibody-based assay for pathogen detection [32]. Compared with the culture-based methods, this immunological approach has been used to detect pathogens in poultry production (poultry feed, feces, litter, carcass rinsing, and water samples) and has provided a better sensitivity and shorter time frame [33]. Recently, improvements by combination with other advanced nanomaterials such as novel enzyme-based signal probes have been made in the basic ELISA method for pathogen detection. For example, by using silica nanoparticles (NPs) carrying poly(acrylic acid) brushes as a “catalase (CAT) container” to increase enzyme loading, Chen et al. [34] presented an improved plasmonic ELISA (pELISA) method for detection of Listeria monocytogenes at ultralow concentrations with the sandwich format. The limit of detection (LOD) obtained by this method (80 CFU/mL) was two and five orders of magnitude lower than that of conventional CAT-based pELISA and horseradish peroxidase (HRP)-based conventional ELISA, respectively. To further simplify the preparation of enzyme-labeled antibody, Lin’s group innovatively prepared an all-in-one organic–inorganic nanoflower, which integrated biorecognition unit (concanavalin A or antibody), signal amplification unit (glucose oxidase or HRP), and carrier unit within a one-pot reaction. And then, it was used for a portable sensitive ELISA detection of Escherichia coli O157:H7. Under the optimal conditions, the detection sensitivity can reach as low as 10 CFU/mL for the case of concanavalin A-glucose oxidase [17] and 60 CFU/mL for the case of antibody-HRP [35].
\nLFIA-based methods are a form of immunoassay, which emerge for the first time at the end of the 1960s and consist of a chromatographic system and immunochemical reaction [36, 37, 38]. The principle of LFIA is based on antibody–antigen specific interaction. After the sample is applied to the sample pad, it migrates along the test strip via capillary action, and a signal response is obtained about 5–10 min later [39, 40]. Due to its simplicity, rapidity, low cost, portability, and facile interpretation without external reagent or external instrumentation, LFIA has held great potential for foodborne pathogen detection [15, 16, 41]. In addition, the LFIA can realize visual detection and quantitative detection by employing different labels, such as colloid gold, fluorescent materials, and magnetic beads [40, 42, 43]. Descriptions on some of the labels that are applied for pathogenic bacteria detection are presented in the following sections.
\nColloid gold is the most widely used label of LFIA due to its intense color and direct visualization [44], and it has been widely used for the detection of foodborne pathogens [45, 46, 47, 48]. Jung et al. [45] used a colloid gold-based LFIA to detect Escherichia coli O157:H7 in enriched samples, and the LOD was 1.8 × 105 CFU/mL without enrichment and 1.8 CFU/mL after enrichment. Preechakasedkit et al. [49] also developed a colloid gold immunochromatographic strip for the detection of Salmonella typhimurium with a minimum detection limit of 1.14 × 105 CFU/mL. Park et al. [15] presented a detection method of Escherichia coli O157:H7 and Salmonella typhimurium with a pressed paper-based dipstick by employing colloid gold as labels. The detection limit of Escherichia coli O157:H7 was around 105 CFU/mL, while that of Salmonella typhimurium was around 106 CFU/mL. In the case of the work of Song et al. [41], Shigella boydii, and Escherichia coli O157:H7 can be detected simultaneously in bread, milk, and jelly samples using colloid gold-based LFIA, and the detection limit of 106 CFU/mL for both Shigella boydii and Escherichia coli O157:H7 was achieved.
\nAs the low sensitivity of colloid gold, fluorescent materials have gained more and more interest due to their higher sensitivity than colloid gold in the field of lateral flow assay [50]. Furthermore, the fluorescent materials enable lateral flow assay to detect the target quantitatively. Compared with colloid gold, which can only provide qualitative or semiquantitative results, quantitative detection can offer more information [42, 51, 52]. In particular, quantum dots show unique fluorescence properties, such as high and stable fluorescence signal [53, 54, 55]. During the last decade, quantum dot-based lateral flow assays have been applied to the detection of foodborne pathogen [56, 57, 58]. Bruno [56] utilized quantum dot-conjugated antibody as the signal reporter of the lateral flow assay to detect Escherichia coli O157:H7. With the assay, the detection limit of Escherichia coli O157:H7 is calculated to be 600 cells per test, while that of colloid gold-based lateral flow assay is 6000 cells per test, indicating the higher sensitivity of quantum dots than colloid gold as labels of lateral flow assay. Chen et al. [58] also developed a competitive format lateral flow assay with quantum dots for the detection of Staphylococcus aureus in food. The detection limit is 3 CFU/mL.
\nMagnetic beads are another type of label, which can realize quantitative detection of targets by measuring the magnetic signal [40, 57, 59]. Due to the fact that they are strongly colored and can enrich and separate targets from complex matrix, magnetic beads are new attractive materials to construct a lateral flow assay, which will probably replace traditional labels. Especially, magnetic beads can simultaneously provide visual signal and magnetic signal. Several researches have recently focused on the use of magnetic bead-based lateral flow assay to detect pathogenic bacteria [60, 61, 62]. Wang et al. [60] employed antibody-coated magnetic beads with the diameter of 300 nm as signal reporter of lateral flow assay for Bacillus anthracis spore detection. A detection limit of 6 × 104 spores/g of milk powder, 2 × 105 spores/g of starch, and 5 × 105 spores/g of baking soda was obtained, respectively. Suaifan et al. [63] described a magnetic bead-based lateral flow assay, which can specifically and simultaneously detect Escherichia coli O157:H7 proteases in complex food matrices. The limits of detection were 12 CFU/mL in broth and 30–300 CFU/mL in food matrices. Xia et al. [64] developed a gold magnetic bifunctional nanobead-based lateral flow assay for the detection of Salmonella choleraesuis. Results indicated that the assay was specific and rapid with the detection limit of 5 × 105 CFU/mL, which was much more sensitive than that of colloid gold-based LFIA (5 × 106 CFU/mL), suggesting that magnetic beads were indeed superior to colloid gold.
\nBesides antibodies, other biomolecules have been investigated to selectively capture and enrich pathogens from cultures, among which aptamer is the most prevalent one [65]. Aptamers, as short single-stranded nucleic acids (DNA or RNA), can bind with high affinity and specificity to a wide range of target molecules, such as ions, small organic molecules, and proteins [66, 67, 68]. The affinities of aptamers for their targets are comparable to, or even higher than most monoclonal antibodies. More importantly, compared with antibodies, they also exhibit a number of advantages. First of all, aptamers can be routinely produced by chemical synthesis, avoiding the use of animals required for antibody production. Furthermore, they are generally more chemically stable, and their binding properties are easier to manipulate. To this end, a number of aptasensors based on optics and electrochemistry have been recently reported for pathogenic microorganism typing and detection.
\nSurface-enhanced Raman scattering (SERS) possesses several attractive properties, such as ultrahigh sensitivity, high speed, comparatively low cost, and multiplexing ability and portability [69, 70, 71], which enable SERS to be widely used for sensitive detection of chemical and biological agents [72, 73]. Since Holt and Cotton first reported the SERS spectrum of bacteria, the identification and detection of microorganism by SERS have attracted high interest recently due to the spectroscopic fingerprint and nondestructive data acquisition in aqueous environment [74]. To date, there have been many SERS biosensors developed, especially based on a “magnetic separation” approach, which focus on bacterial pathogen detection. Wang et al. [75] reported a magnetically assisted SERS biosensor for single-cell detection of Staphylococcus aureus on the basis of aptamer recognition. The biosensor consists of two basic elements including a SERS substrate (Ag-coated magnetic nanoparticles) and a novel SERS tag (Au nanorod-5,5-dithiobis-(2-nitrobenzoic acid) (AuNR-DTNB)@Ag-DTNB core-shell plasmonic NPs or DTNB-labeled inside-and-outside plasmonic NPs (DioPNPs)). Based on these, the LOD of 10 cells/mL can be achieved for Staphylococcus aureus detection. Similarly, through combined gold NPs (GNPs) modified with Raman molecules and Fe3O4 magnetic GNPs immobilized with aptamer, Zhang et al. [76] successfully fabricated GNP-enhanced SERS aptasensor for the simultaneous detection of Salmonella typhimurium and Staphylococcus aureus. In comparison with these label-based SERS methods, label-free methods do not require a secondary label dye and can directly obtain the intrinsic fingerprint of bacteria, which relies on the mutual interaction of bacteria cell with the SERS substrate [77]. With this regard, a lot of label-free methods have been developed for the detection of pathogens. For example, Gao’s group [78] successfully achieved intuitive label-free SERS detection of bacteria using aptamer-based in situ Ag NP synthesis. The biosensor as prepared can recognize bacteria quickly and directly by SERS with the formation of well-defined bacteria-aptamer@Ag NPs. The detection limit is down to 1.5 CFU/mL.
\nAs another typical spectroscopic method, fluorescence resonance energy transfer (FRET, a homogeneous signal transduction technique), has been gradually employed for the determination of pathogenic bacteria. Yu et al. [79] presented a universal and facile one-step strategy for sensitive and selective detection of pathogenic bacteria using a dual-molecular affinity-based FRET platform based on the recognition of bacterial cell walls by antibiotic and aptamer molecules, respectively. Within 30 min, the FRET signal shows a linear variation with the concentration of Staphylococcus aureus in the range from 20 to 108 CFU/mL with a detection limit of 10 CFU/mL. Moreover, Duan’s group [80] further achieved simultaneous detection of Vibrio parahaemolyticus, Staphylococcus aureus, and Salmonella typhimurium through using multicolor dyes as donors and carbon NPs as a sole acceptor in FRET.
\nCompared with optical-based biosensors, electrochemical methods, in general, show the potential for construction of fast, simple, low-cost, sensitive, and high-throughput biosensors that can be miniaturized [81, 82, 83, 84]. To date, electrochemical aptasensors are widely used for identification and quantification of pathogens. For example, Labib et al. [85] developed an impedimetric sensor via assembling their selected highly specific DNA aptamers onto a gold NP-modified screen-printed carbon electrode for the highly sensitive detection of live Salmonella typhimurium. This aptasensor is very simple and highly selective. It can successfully detect Salmonella typhimurium down to 600 CFU/mL (equivalent to 18 live cells in 30 μL of assay volume). Moreover, to further improve the sensitivity, Abbaspour et al. [86] innovatively combined the magnetic beads’ fast separation with the Ag NPs’ signal amplification. They successfully fabricated an electrochemical dual-aptamer-based sandwich detection method for Staphylococcus aureus. The aptasensor as prepared shows an extended dynamic range from 10 to 1 × 106 CFU/mL with a low detection limit of 1.0 CFU/mL (S/N = 3). Despite much progress has been made, these methods always require probe labeling and aptamer immobilization, which may affect the binding affinities between bacteria and their aptamers. With this respect, Ding’s group [87] constructed a label-free potentiometric aptasensor for rapid, sensitive, and selective detection of Listeria monocytogenes. In this strategy, the target-binding event prevents the aptamer from electrostatically interacting with protamine, which can be sensitively detected using a polycation-sensitive membrane electrode.
\nCulture-based foodborne pathogen detection methods, although sensitive enough, are often too time-consuming to reduce foodborne disease occurrence. Therefore, a large number of innovative methods have been developed to overcome this performance limitation. These rapid detection methods can be classified into nucleic acid-based methods, antibody-based methods, and aptamer-based methods. All these rapid methods for foodborne pathogen detection are superior to culture-based methods. However, some of them still require improvement in sensitivity, selectivity, simplicity, or accuracy to be of any practical use. Nucleic acid-based methods, as a replacement method for culture-based methods, have high sensitivity and require a shorter time than conventional culture-based techniques for foodborne pathogen detection. Most of them still require highly trained personnel and expensive instruments, which limit their use in a practical environment. The development of antibody-based methods helped improve the time required to yield results. The specific binding of antibody to its antigen results in its high specificity and sensitivity of antibody-based methods, and they work well in food matrices without being interfered by other DNAs, proteins, or nontarget cells. Aptamer-based methods are similar to antibody-based methods, which also exhibit high sensitivity and selectivity. However, they still need to be improved for food matrix detection. Increasing detection accuracy and decreasing detection time are the eternal themes in rapid detection. In the future, new nanomaterials and rational biosensing strategies would be developed to approach the goal.
\nSelf-limited epilepsy with centrotemporal spikes (SECTS), well-known as Rolandic epilepsy is the most frequent among the childhood focal epilepsies and may account for about 15–25% of all epileptic syndromes diagnosed between the ages of 5 to 15 years [1]. It is termed ‘rolandic’ epilepsy because the focal seizures are originated from the region around the lower part of the central gyrus of Rolando. The incidence range changes between 7.1–21 per 100000 in a population younger than 15 years with male predominance [2]. The age of onset in 90% of cases between 1 and 10 years with a peak around 6–7 years and recovery occurs before the age of 15–16 years [2, 3, 4].
Self-limited epilepsy with centrotemporal spikes is a syndrome of brief hemifacial motor seizures, frequently having associated somatosensory symptoms, usually without impairment of consciousness which tend to evolve into GTCS [3, 4, 5, 6]. Seizures are often related to sleep [7]. Genetic predisposition is frequent, and there is male predominance [3, 5, 8, 9]. An interictal EEG has normal background activity with biphasic high-voltage centrotemporal spikes, often followed by slow waves that are activated by sleep and tend to spread or shift from side to side [10]. Neurological and mental status before the debut of epilepsy is normal. There are no specific abnormalities on brain MRI or CT. Many studies reported that RE may cause transient or long-lasting cognitive and behavioral disturbances [4, 5, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40].
Panayiotopoulos described a concept of benign childhood susceptibility syndrome (BCSSS) to unify RE, Panayiotopoulos syndrome (PS) and childhood occipital epilepsy of Gastaut (ICOE-G) outlined the common features, course of diseases, prognosis, and the possible genetic predisposition in this group of associated syndromes [5].
1989 ILAE classification recognized three “age-related and localization-related epilepsies and syndromes”: (1) benign childhood epilepsy with centrotemporal spikes (BCECTS); (2) childhood epilepsy with occipital paroxysms; (3) primary reading epilepsy [10].
ILAE Commission on Classification and Terminology lists three childhood idiopathic focal epilepsy syndromes: (1) benign childhood epilepsy with centrotemporal spikes (BCECTS); (2) Panayiotopoulos syndrome, and (3) late-onset childhood occipital epilepsy (Gastaut type) [13].
Rolandic epilepsy had undergone significant terminological and classification changes. RE had been previously considered as benign childhood epilepsy. Frequently reported cognitive and language impairments and behavioral disturbances in children with RE led to the replacement of the terms “benign” and “idiopathic” by the “self-limited” in the new classification proposed by Sheffer I. and colleagues [41].
The role of genetic factors in RE has been presumed since the first high incidence of centrotemporal spikes in family members of patients with RE was reported in 1964 [42]. RE and related syndromes with atypical features do not follow a Mendelian inheritance mode [43]. The clinical and genetic studies have shown complex inheritance [43, 44, 45, 46, 47, 48, 49, 50, 51].
The genetics of CTS is not the same as the clinical genetics of RE [52]. Although CTS is the primary EEG characteristics of RE or ARE, they are also observed in healthy children [53] or the children with autistic spectrum disorders without seizures [54]. Only 10% of EEG trait carriers had seizures [55, 56]. An autosomal dominant mode of inheritance of CTS on EEG has been reported by several authors [42, 55, 56] but it is still debated [52]. The linkage of CTS to ELP4-PAX6 region on 11p13 and chromosome 15q13 [57], 16p12–11.2 [58], and 15q14 [59] have been identified.
Doose et al. investigated the broad spectrum clinical and EEG manifestation of 147 children with RE and their 1266 family members revealed a high incidence of febrile convulsion and afebrile GTCS in patients and their relatives suggested multifactorial inheritance. EEG recordings of probands and their siblings showed a high rate of generalized EEG traits [43].
A multicentral twin study of eighteen twin pairs (10 MZ, 8 DZ) based on a twin database done by Vadlamudi and colleagues demonstrated that the etiology of RE and its inheritance mode is much more complicated than considered before [44].No twin pairs were concordant for RE. Only one monozygotic twin pair has shown centrotemporal spikes on EEG without seizures. Another intriguing finding from this twin data was that all twin pairs with atypical features RE, had a co-twin with seizures although discordant for RE, which emphasized that genetic factors may be more important in atypical cases of RE.
The genetic basis of RE/ARE is polygenic and complex, the interaction of environmental factors or other genes should be considered in etiology of RE spectrum epilepsy syndromes [60, 61]. A number of genes were found to follow the Mendelian inheritance and be associated with RE/ARE (Table 1).
Mutations | Protein function | Special features | Reports |
---|---|---|---|
KCNQ2/KCNQ3 | Voltage-gated potassium channel | BFNS plus RE | Maihara et al. [62] |
BFNS CTS trait | Coppola et al. [63] | ||
BFNS plus RE, RE, CTS trait | Neubauer et al. [64] | ||
SRPX2 | E2A/HLF fusion | RS, oral and speech dyspraxia, MR | Roll et al. [65] |
ELP4 | Elongator subunit | CTS trait, speech disorder, behavior disturbances, ADHD | Strug et al. [57] |
GRIN2A | NMDAR subunit | Atypical RE (LKS/CSWS), intellectual disability, various dysmorphic features | Lemke et al. [66] |
RBFXO1/3 | ATAXIN 2-BINDING PROTEIN 1/HEXARIBONUCLEOTIDE-BINDING PROTEIN 3 | RE, atypical RE, CTS trait | Lal et al. [67] |
DEPDC5 | GATOR complex | RE, atypical RE | Lal et al. [50] |
GABRG2 | GABA receptors | RE, atypical RE | Reinthaler et al. [68] |
CAMK2A | Subunit of calcium/calmodulin-dependent protein kinase II | Atypical RE, intellectual disability and autism | Rudolf et al. [45] |
GRIN2B | NMDAR subunit | Atypical RE, epileptic encephalopathy, intellectual disability | Rudolf et al. [45] |
CHRNA4 | Neuronal nicotinic acetylcholine receptor α4 subunits | RE (familial case) | Neng et al. [69] |
Genetic mutations associated with RE/ARE spectrum.
Lemke et al., have identified GRIN2A mutations in 20% of patients with ARE associated with neurocognitive disturbances [66].
Although mutations in PRRT2, KCNQ2, KCNQ3, RBFOX1, and DEPDC5 genes with an autosomal dominant transmission reported in patients with RE spectrum epilepsy syndromes, they have not been confirmed by the studies based on large case series [70].
With the exception of GRIN2A and ELP4, many genes currently associated with RE/ARE, including KCNQ2, KCNQ3, CHRNA4, DEPDC5, RBFOX1/3, BDNF, and GABAA-R, were initially linked to other neurogenetic conditions, and later their phenotypes were expanded to RE/ARE.
The main seizure type in RE according to the ILAE 2017 seizure classification is focal aware seizure consisting of motor-hemifacial tonic or clonic contractions, oro-pharyngo-laryngeal symptoms, sensory symptoms represented by unilateral numbness or paresthesia of tongue, lips, gum, and inner part of the check, and associated with speech arrest, hypersalivation, and focal to bilateral seizures [2, 3, 4, 41, 71]. Hemiconvulsions and bilateral tonic–clonic seizures are less frequently observed ictal features, mainly seen in younger children due to rapid distribution of focal onset seizures [2, 3, 4, 6]. Hemiconvulsions may be followed by post-ictal Todd’s hemiparesis in 10% of cases [8, 72].
Seizures are brief, usually last from 30 sec to 2–3 minutes or longer if turn into bilateral tonic–clonic seizures [6, 18, 72]. Seizures mainly occur during night sleep or drowsiness, whereas the probability of awake seizures is less than 10% [73, 74]. Seizure frequency is low, most patients have less than 10 seizures, 10%–20% of patients have a single seizure [75]. Consciousness is completely preserved in around 60% of patients with RE [5].
Focal motor seizures in approximately one-third of cases manifest as unilateral oral-facial tonic or clonic contractions. These are brief (few seconds −1 min), a sudden burst of clonic contractions of the face, which may be entirely localized in the lower lip or spread to the ipsilateral upper and very rare to the lower extremities [1, 2, 3, 4, 5, 71, 76].
Tonic deviation of the mouth is frequently observed ictal motor manifestation [5].
Oro-pharyngo-laryngeal symptoms are mostly motor ictal phenomena with the involvement of the (epi-) glottis and pharynx (> 50%) produce guttural bizarre sounds, resembling gargling, grunting, wheezing [72]. These may be accompanied by contractions of the respiratory and abdominal muscles (vomiting like contractions) which appear in more than half of seizures [77]. They consist of unilateral sensory and motor manifestations inside the mouth, tongue, inner cheek, gums, teeth, and pharyngolaryngeal regions [3].
Speech arrest occurs in >40% of seizures with dys - or anarthria [3, 72]. The child usually is aware, with preserved receptive language, attempts to communicate with gestures, but unable to produce a single intelligible word [3, 5]. Speech arrest is considered more as a motor ictal manifestation associated with the loss of the power and coordination for the articulation of words [3]. There is no impairment of the cortical language mechanisms [4, 5].
Focal non-motor seizures commonly observed in RE.
Sensory symptoms may manifest as unilateral numbness or paraesthesias like tingling, prickling, freezing and their variations in the parts (rarely involve the whole area) of oral-facial-pharyngeal area, usually tongue, inner cheek, gum, teeth, lips [3, 4, 6]. Sensory seizures often occur in combination with motor seizures and hypersalivation [3, 5, 72].
Hypersalivation is one of the most characteristic autonomic ictal symptoms of RE, occurs in one-third of cases [2, 3, 4, 5, 71]. It is frequently associated with hemifacial motor symptoms. As well as the awareness is not disturbed in most of the cases, children usually are able to describe their sensations as sudden filling of the mouth with saliva and air, difficulty in pronouncing words, a lot of saliva flowing from the mouth [5].
Other autonomic ictal manifestations as ictal emesis and ictal syncope may observe rarely in RE. Although autonomic seizures are the cardinal symptom of Panayiotopoulos syndrome, they are reported in RE [74, 78, 79, 80, 81, 82]. The overlap of the clinical and EEG features of PS and RE has been widely investigated by several authors [5, 74, 79, 80]. The cases where two different types of childhood focal seizures presented at the same time or one form of epilepsy progressed to another have been thoroughly reported by different investigators [74, 79, 80, 81, 82, 83, 84, 85].
Focal to bilateral tonic–clonic seizures are a frequent seizure type present in one to two-thirds of children with RE. FBTCSs mostly appear during night sleep [86].
Status epilepticus is seen rarely and usually associated with an atypical course of the disease [87].
Focal motor SE occurs more often than generalized convulsive SE [3]. This state consists of unilateral or bilateral hemifacial contraction, subtle perioral myoclonus, speech arrest, dysarthria, excessive drooling, swallowing difficulties [88, 89, 90, 91, 92, 93].
The EEG picture is distinctive in Rolandic epilepsy. The background activity is almost always preserved in an awake state and during sleep [91]. The characteristic interictal EEG pattern- centrotemporal spikes (CTS) or rolandic spikes are regarded as the neurobiological markers of RE. CTS is high-amplitude (usually > than 150 mkV) biphasic spikes or sharp waves of ∼ 70–80 milliseconds duration frequently followed by a slow activity on the central-mid temporal region (C3/C4, T3/T4) [2, 3, 4]. More posterior localization of CTS is often observed in the youngest patients [94]. The spikes may occur isolated or in clusters, in one or both hemispheres [95, 96] (Figure 1A and B). A focal rhythmic slow activity over the centrotemporal region is occasionally observed [2]. The most typical finding of the rolandic spikes is their significant increase in frequency during NREM sleep [74] (Figure 1C-E). The spikes appear only in sleep in about a third of children [97].
7y.old boy with rolandic epilepsy. (A) Interictal awake EEG. Left centrotemporal spikes with rapid spreading to the right frontal-anterior temporal region. with maximum negativity over the C3T3,F7 and maximum positivity on the right-midline frontal-parietal region F4Fp2FzPzCz(AV montage, sensitivity 20Mkv/mm, paper speed 30mm/sec, LFF 70 Hz, HFF 0.5 Hz, Rejector 50 Hz). (B) Increased paper speed clearly demonstrates propagation of left centrotemporal spikes to the right frontal-anterior temporal region (AV montage, sensitivity 20 Mkv/mm, paper speed 120 mm/sec, LFF 70 Hz, HFF 0.5 Hz, Rejector 50 Hz). (C and D) Interictal 1NREM sleep EEG. The frequency of CTS of the same distribution is increased. (E) The same EEG sample in bipolar longitudinal montage demonstrates phase reversal on both right and left centrotemporal region (C3C4T3T4).
EEG and magnetoencephalography (MEG) studies show a stable horizontal dipole coming from the lower rolandic region with maximal electronegativity in the centrotemporal region and electropositivity in the frontal region, usually seen unilateral or bilateral [98, 99, 100]. Spikes may often appear in the central, parietal, midline, or even occipital regions which do not exclude a diagnosis of RE [98].
Somatosensory stimulation by the tapping of hands or feet or electrical stimulation of fingers at 1 Hz may activate CTS and somatosensory evoked potentials (SEPs) on the contralateral hemisphere [101].
Brief interictal generalized bursts of 3–5 Hz slow waves with intermixed small spikes distinctive than a pattern 3 Hz spike–wave seen in CAE may observe in about 4% of patients with RE [85, 102].
Many studies have tried to identify the source of rolandic discharges using topographic analysis, source modeling techniques, dipole tracing method, magnetoencephalography (MEG), and functional MRI (fMRI) investigations [99, 100, 101, 103, 104, 105, 106]. The functional MRI (fMRI) triggered by EEG of the rolandic spikes as well as MEG showed activation of the sensorimotor area [104], mainly in the orofacial division of the primary sensorimotor cortex [105]. However, it is challenging to distinguish the precentral or postcentral origin of CTS [103]. Ishitobi et al., suggested the precentral origin of rolandic spikes explained this theory by the continuity of cortical surface polarity from negative gyral cortex to the surface positive interhemispheric fissure based on the combination of scalp EEG and MEG [103]. Gregory and Wong analyzed 12 independent foci in 10 patients with RE assumed that the generator of a dipole discharge was located halfway between the maximum negative and positive poles, and was most likely situated at the depth of the lower rolandic fissure or Sylvian fissure [107]. The propagation pattern of rolandic spikes first studied by Jung et al., suggests that spike propagation was caused by intracortical spreading a single dipole across the central sulcus [108].
CTS are diagnostic markers of RE only in a suggestive clinical presentation [74]. It has been widely reported that 1.2 to 3.5% of normal healthy children population between 5 and 12 years old [109, 110], 6–34% of siblings and relatives of patients affected by RE [9, 111, 112], children with migraine, behavior disturbances, ADHD, variety of organic brain diseases with or without seizures, such as cerebral tumors, Rett syndrome, fragile X syndrome and focal cortical dysplasia [84, 113] also show CTS in routine EEG recording.
The first described ictal patterns are characterized by a quite monomorphic sequence of rhythmic sharp waves or spikes without significant post-ictal slowing [91, 114]. In 1990, Gutierrez et al. described an ictal event with speech arrest only characterized by a short train of ictal alpha activity, and then two multiple spikes and wave complexes originated from the left centrotemporal region followed by marked attenuation of the left hemispheric background [115]. Subclinical rhythmic discharges of spike and wave in the centrotemporal region have been documented by several authors in RE [116, 117]. Saint-Martin et al. in 2001 described a series of patients presenting with typical and also atypical ictal manifestations such as falls, negative myoclonus and observed that positive motor phenomenon correlated to the spike component preceding a negative motor phenomenon, correlated with the slow-wave component of the spike and wave complex [118].
Capovilla et al. recorded 34 seizures in 30 patients with RE and described four electrographic seizure patterns thus emphasizing that ictal pattern for RE is not unique [116]:
low-voltage activity of fast rhythmic spikes, increasing in amplitude and decreasing in frequency observed in the majority of patients,
a discharge of spikes intermixed with sharp waves increasing in frequency and amplitude,
monomorphic theta which progressively formed a discharge increasing in amplitude and decreasing in frequency,
initial focal depression of the electrical activity, followed by one of the three above described patterns.
Ictal EEG source analysis of 3 patients with RE demonstrated the activation of the opercula-insular area, time-locked to the contralateral focal myoclonic jerks [119].
In most cases, children with RE have a good prognosis regarding both seizures and neurodevelopment [120, 121]. The remission of seizures usually occurs before the age of 18 years [11, 98]. The cognitive and behavior problem may happen in an active period of disease which is reversible in most patients [11, 12].
Rolandic seizures occur in a period of significant brain maturation. The dysfunction of neuronal network activities such as focal discharges may be associated with neuropsychological problems, including, linguistic, cognitive, and behavioral impairment [28, 29, 30, 122]. The frequent spike discharges in sleep may boost language and attention processing problems [120, 123, 124, 125].
Mood and behavioral disorders were present in nearly a third of children (30.9%) with RE [126, 127, 128]. Retrospective studies have proposed that early age at onset pretends a more aggressive seizure course [18, 129, 130, 131].
Functional MRI study revealed CTS density caused hemodynamic changes even during wakefulness can interfere with the normal brain-language network and the bilateral insular cortex [132].
The neuropsychological tests such as Wechsler Intelligence Scale for Children-III (WISC-3rd), verbal fluency test, Wisconsin card sorting test, attention deficit diagnostic scale, and child behavior checklist scale are usually administered to measure a wide range of skills and cognitive functions of RE patients [35, 36, 37, 38, 133].
Many researchers showed a variety of neuropsychological deficits, behavioral and emotional difficulties in a limited cohort of patients with RE range from 19 to 67% [22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 134, 135]. The series of reported children with uncomplicated RE were described lower average results on neuropsychological tests involving visuomotor coordination, some executive functions, sustained attention, and language issues like spelling, reading aloud, reading comprehension, memory, and learning of auditory–verbal material, delayed recall, and verbal fluency, compared with controls. However, the Full-Scale Intelligence Quotient (FSIQ) was not significantly low in most of them.
D’Alessandro et al. investigated the neuropsychological data of 44 children with RE who did not have a seizure for more than 6 months without treatment. Attention, language, and visuomotor coordination tasks problems were more severe in children with a bilateral epileptiform discharge. However, in a follow-up assessment for 4 years, a re-examination of 11 children had revealed the normalization of cognitive functions in all [21].
Several studies reported that cognitive abnormalities and behavioral impairments are associated with a high interictal spike frequency [24, 25, 35, 120], the number of interictal abnormalities in wake or sleep [136], activation of interictal spikes during sleep [118, 137], and the presence of non-tangential dipole spikes [73].
Piccinelli et al. [138], investigated the frequency of specific learning disabilities such as reading, writing, and calculation in patients with typical RE and possible related electroclinical findings. They reported children with RE who developed seizures before age 8 years and had epileptiform discharges more than 50% of the sleep EEG recording in several tracings over more than a year were at risk of developing academic difficulties [138].
EEG may predict educational and behavioral impairments in children with RE. The presence of an intermittent slow-wave focus during wakefulness, a high number of spikes in the first hour of sleep (and during whole night sleep), and multiple asynchronous bilateral spike–wave foci in the first hour of sleep are associated with learning problems in children with RE [16, 139].
RE can present or evolve to an atypical form, characterized by atypical ictal semiology, different EEG findings, and poor neuropsychological outcomes [19, 140, 141].
Massa et al. described 5 interictal EEG patterns that significantly correlated with atypical evolutions of RE: [41] intermittent slow-wave focus; [2] multiple asynchronous spike–wave foci; [3] long spike–wave clusters; [4] generalized 3-c/s “absence-like” spike–wave discharges; [1] conjunction of interictal paroxysms with negative or positive myoclonia, and abundance of interictal abnormalities during wakefulness and sleep [136].
Several studies have shown an association between atypical rolandic epilepsy and known genes (Table 1). The identification of de novo or inherited mutations of N-methyl-D-aspartate (NMDA) receptor subunit-encoding genes (GRIN2A and GRIN2B) linked to speech and language, cognitive impairment, and behavioral difficulties have been a significant breakthrough in the understanding of the nature of atypical RE [142, 143, 144, 145]. Another relevant gene is elongation factor protein 4 (ELP4), which is associated with language impairment, autism spectrum disorder, mental retardation, and epilepsy with centrotemporal spikes on EEG [146].
Atypical rolandic epilepsy (ARE) is a severe epileptic condition especially with regards to cognitive consequences. The first description of atypical features of RE was published by Aicardi & Chevrie in 1982 showed rolandic epilepsy presenting periods with new types of seizures, mainly atonic and myoclonic, associated with continuous spike-and-waves in slow-sleep EEG (CSWS/ESES), and transitory learning difficulties [147]. Doose and Baier described similar patients with atonic fits leading to daily falls which is the hallmark seizure type for Lennox–Gastaut syndrome and termed the condition “pseudo-Lennox syndrome” to differentiate this two distinct conditions [148]. Patients with ARE have significantly lower full-scale and verbal IQ than the patients with typical RE [149]. Neuropsychological impairment, which may sometimes be present before the onset of the disease, is constantly present during the clinical course, but in contrast to ESES and LKS, the cognitive outcome is always favorable [92, 150]. Clinical semiology consists of typical for RE focal seizures, generalized tonic–clonic seizures, atypical absences, myoclonic seizures, and atonic seizures. The atonic attacks may involve the whole axial musculature or be localized, causing repeated brief (0.5–2.0 s) atonic episodes in the head or a limb (epileptic negative myoclonus) that usually occur for periods lasting one to several weeks, separated by seizure-free intervals of weeks or months [6, 90, 92]. Such atonic attacks are associated with the slow-wave component of spike and wave complexes, and the location of the EEG discharges corresponds to that of the atonic episodes [151, 152]. Interictal awake EEG shows bilateral sharp and sharp-slow wave complexes with higher amplitude in the rolandic area, which increases during sleep with bilateral synchronization [90, 92, 116, 153].
Using carbamazepine may promote the diffusion of spike–wave activity from the rolandic focus to induce atonic seizures, atypical absences in patients with RE [154].
Rolandic status epilepticus refers to status epilepticus that can be convulsive or non-convulsive, and either generalized or focal lasting days or weeks including motor facial seizures, oromotor dyspraxia, anarthria with persistent drooling and swallowing problems [155]. The interictal EEG usually shows focally or bilaterally synchronous sharp waves or sharp and slow wave complexes predominant in the rolandic area with a tendency to become continuous during sleep [146, 155]. The condition can be resolved with a good neurocognitive outcome with appropriate treatment [146]. These seizures can persist for more than 1 month without treatment [156, 157].
The decision whether to treat children with RE or not requires a particularly careful risk–benefit analysis [2, 158, 159, 160, 161, 162, 163]. Many authors suggest that drug treatment is not necessary for typical RE because of its good prognosis, and usually infrequent nocturnal seizures [114, 154]. Moreover, in 40–50% of cases, the seizures are difficult to control with drugs [148]. Besides, the treatment with AED usually does not influence the duration of active epilepsy [163].
However, treatment may be indicated in patients with frequently recurring daytime seizures, generalized tonic–clonic seizures, young age at onset [164], or when the ictal events are disruptive to the patient or family [161, 163]. Furthermore, the presence of cognitive and behavioral disturbances, either transitory or persistent has to be considered [2, 5, 91, 92]. There is no single solution supported by definitive evidence which AED is more effective in the treatment of RE.
Internationally, carbamazepine (CBZ 20-40 mg/kg/d [165]) and valproate (VPA 20-30 mg/kg/d [166]) are the most often prescribed AED for children newly diagnosed RE [167]. However, the possible worsening of EEG in rolandic epilepsy by some drugs and particularly by CBZ, increasing epileptiform abnormalities during sleep, and inducing epileptic negative myoclonus have been reported [154].
Sulthiame, levetiracetam, and gabapentin were studied in a randomized controlled trial [158, 159, 167, 168, 169]. Sulthiame administered varied between 3.1 and 5.7 mg/kg/day was effective in controlling seizures in children with RE [159].
A prospective, open-label, pilot trial evaluating the efficacy and tolerability of levetiracetam (LVT 20-30 mg/kg/d) or oxcarbazepine (OXC 20-35 mg/kg/d) as monotherapy in two parallel groups of newly diagnosed RE patients demonstrated effectiveness in controlling seizures a follow-up period up to 2 years [162].
A randomized controlled multicenter trial comparing the effects of either Levetiracetam or Sulthiame on EEG in RE showed a reduction of epileptiform discharges after 12 weeks of treatment [158]. Persistent epileptiform discharges after 12 weeks of treatment are associated with recurrent seizures [158].
When the presence of ESES associated or not with negative myoclonus, clinical status, or acquired aphasia is detected in children with RE, a change of antiepileptic drugs should be considered. Class IV studies suggest that sulthiame, benzodiazepines, ethosuximide, and, in most severe cases, corticosteroids might be useful [91, 92].
Duration of treatment in RE should not exceed 1 year following the last seizure, regardless of EEG changes [2].
We are grateful to Dr. Sándor Beniczky for his valuable comments on EEG figures.
None of the authors has any conflict of interest to disclose.
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