Sexually transmitted diseases (STDs) are among the most common infections. Their clinical identification is difficult because STDs are often asymptomatic. Untreated infections with these pathogens can in time lead to serious consequences. It is documented that isolation of some of these bacteria from cultures is very difficult. Because there is a large number of STD pathogens which can generate coinfections, their simultaneous detection in a unique sample is very important. Multiplex polymerase chain reaction (PCR) is an advanced method of molecular biology which allows for simultaneous detection of multiple pathogens in the same sample. The advantages of the multiplex PCR method were assessed by various researchers by comparing the diagnosis results obtained with different other conventional methods. The sensitivity and specificity of these methods were analyzed on different specimens in comparison to traditional methods, such as culture media or direct microscopic examination. These studies demonstrated beyond any doubt that the multiplex PCR system is highly effective in the detection of each of multiple STD pathogens depicted from a single specimen and argued for multiplex PCR superiority in terms of sensitivity and rapidity.
- STD pathogens
- simultaneous detection
- multiplex PCR
Sexually transmitted diseases (STDs) are an issue of great interest as they are among the most common of all infections. It is documented that STDs are more prevalent in economically underdeveloped populations. Only a few countries outside Western Europe and North America have implemented monitoring systems for these infections. In the rest of the world, epidemiological studies are based on the results of samples provided by certain population segments (mainly symptomatic patients or prenatal controls) not necessarily representative for the majority of the population. Usually, persons presenting obvious signs (secretions, lesions or pain) interrupt their sexual activity and seek medical help. Ironically, most often those who actually transmit these infections are asymptomatic or present mild symptoms .
It is well‐known that vaginal bacterial infections can induce human infertility, yet they are underestimated in infertility testing. Population screening and treatment of individuals presenting unexplained infertility that are detected as STD‐infected persons seem appropriate in this context .
No STD can be regarded as an isolated problem since multiple infections are rather common and STD infections generally denote a high‐risk sexual behavior that often can be associated with more serious infections. It is therefore important to acknowledge that STDs are often asymptomatic or cause nonspecific symptoms and that periodical STD testing is crucial in limiting the risk of human immunodeficiency virus (HIV) infection. STD control could significantly reduce HIV incidence worldwide, although the impact of interventions may vary depending on local epidemiological contexts. Analysis of data from several studies has suggested that a better management of STD cases is more likely to reduce HIV incidence in the early stages of an epidemic, when HIV infections are concentrated in population groups with a high prevalence of other curable STDs .
A correct STD diagnosis is needed to prevent further spreading of such infections in the healthy population. Several techniques and laboratory methods for highlighting these diseases were developed in the past decades. For instance, because of their high sensitivity, specificity and suitability for different types of samples, nucleic acid amplification tests (NAATs) are suitable for the diagnosis of urogenital infections. Lately, however, polymerase chain reaction (PCR) techniques are increasingly employed in such cases as they allow direct, sensitive, automated and usable detection of STD‐causing pathogens on all sample types and even the simultaneous detection of several STDs.
2. The most frequent causative agent of STDs
Urogenital infection with
Several data indicate the involvement of the gonococci in miscarriages. Screening and medical management of the
The term mycoplasma is used to designate organisms in the Mollicutes class, the smallest free‐living cell‐wall‐deficient microorganisms, the most simple life forms capable of replication outside a host cell.
Several authors have reported that these organisms are engaged in women infertility, preterm delivery, premature rupture of membranes and chorioamnionitis [24, 25]. In addition, some studies present mycoplasma as causative agents of male infertility. It has been reported that these infections change various sperm characteristics, such as motility, density or morphology, and that antibiotic treatment improves the quality of the sperm . A recent study found out that detection rates of
3. STD detection methods
Due to technical difficulties resulting from the need to inoculate specimens immediately after sampling, culture assays present low sensitivity (50%) and are seldom used today in the microbiological detection of
It is thought that
Today, there are numerous methods available for
Precise diagnosis of gonorrhea is needed to prevent severe complications and to control transmission, especially in the case of asymptomatic infections. Molecular approaches such as hybridization assays or nucleic acid amplification tests have revolutionized the diagnosis of gonococcal infection due to their increased accuracy compared to the culture media and their ability to simultaneously test multiple species. NAAT should be the technique of choice in the diagnosis of coinfections and screening. In addition to diagnostic, molecular approaches have been successfully applied for testing
Microscopic examination of the smear and/or culture is presently the most commonly used method in the detection of
The contribution of flow cytometry technology in investigating adhesion of the extracellular parasite to human host cells has been clearly demonstrated. This methodology can be optimized and the test can be used in a format in which several different strains may be analyzed simultaneously . It is, however, a difficult method for the diagnosis of trichomooasis. A recent study  indicated the advantages of using DNA in the detection of
As mentioned above,
The safe detection of
Over 50 mycoplasma genome sequences are now available in public databases, revealing a genetic diversity more complex than first predicted. This growing set of data is extremely valuable in the study of organisms otherwise difficult to cultivate, offering new means for testing and molecular diagnostics .
4. Multiplex PCR methods as diagnostic tools for STDs
Multiplex PCR methods for the simultaneous diagnosis of several STDs are more and more often employed in recent years. These methods place a number of primer pairs corresponding to specific DNA sequences of various STD pathogens all in one reaction tube, such commercial ready to use kits being available. While some kits can simultaneously detect two commonly associated STDs, such as
Another asset is that easily collectable urine samples are adequate DNA sources for these PCR techniques along with the urethral/vaginal discharge swabs. About 30–50 mL of first void urine (FVU) samples collected in sterile polypropylene containers early in the morning, at least 4 h after the previous urination, are more than enough for a successful analysis. FVU is definitely less invasive compared to the harvesting of vaginal or urethral secretions, an aspect reported to have influenced a significant number of subjects to avoid STD identification tests previously . As urine specimens can be self‐collected in intimacy, an increased number of patients tend to favor such option that also enhances the applicability of the screening programs . The fact that urethral secretions were found to present increased sensitivity and specificity compared to FVU in immunological analyzes for male subjects seems to hold lesser relevance, anyway differences have diminished since NAAT were first performed . Concerning the female subjects, the use of urine specimens in STD identification produced similar results to vaginal or endocervical secretions . It is known that assay sensitivity may decrease if samples are repeatedly frozen/thawed or stored for longer periods of time because nucleic acids can easily degrade. Urine samples hold another advantage as well: they can be stored at 4–8°C up to 7 days prior to the processing.
Thanks to this easy and noninvasive sampling, the multiplex PCR can be used to detect STDs in both symptomatic and asymptomatic individuals and could prove to be a useful screening tool for the general population.
A 15 min centrifugation at 15,000g and subsequent supernatant removal and pellet resuspension are needed as a successful DNA extraction requires concentrated pathogen suspensions. Both urethral/vaginal discharge swab specimens and urine samples should be brought to room temperature prior to the centrifugation. One can use up to 10 mL amounts of sample to increase the DNA extraction yield. Several commercially available kits for DNA extraction can be used according to the manufacturer's instructions, e.g., QIAamp® DNA Mini Kit (Qiagen, Hilden, Germany), AccuPrep Genomic DNA Extraction Kit (Bioneer, Seoul, Korea) or MasterPure™ Complete DNA and RNA Purification Kits (Epicentre Biotechnologies, USA).
In our marketing era, such new techniques and gadgets (kits) need to be accompanied by studies assessing their performance compared to other traditional methods.
One such study conducted in 2010  compared the multiplex PCR assay for the simultaneous detection of
Another study  in 2011 compared the results of 113 STD patients tested for six sexually transmitted microorganisms (
Seeplex® STD6 ACE Detection kit (Seegene, Korea) was evaluated in yet another Korean study, completed in 2012, which was conducted on 739 subjects . Cervical swabs were collected from the women enrolled, while men were requested to provide 30–40 mL of FVU. The six pairs of dual priming oligonucleotide primers specifically targeting the
Anyplex™ II STI‐7 Detection Kit (Seegene, Korea) was tested to the limit in a 2013 Korean study  aiming to investigate the accuracy and reliability of a real‐time multiplex PCR assay employed for the detection of seven STD agents (
We ourselves have used a Seeplex® STD6 ACE Detection kit in a study on 224 Romanian subjects including persons with STD symptoms and asymptomatic individuals involved in unprotected sex with multiple partners, couples experiencing unexplained infertility or others requesting STD screening for various reasons. Each individual contributed with 30–50 mL FVU samples and DNA extraction was accomplished with MasterPure™ Complete DNA and RNA Purification Kits (Epicentre Biotechnologies, USA). A 2% agarose gel containing ethidium bromide was used with 0.5× Tris‐borate 0.1 mM EDTA (TBE) as running buffer. The PCR products were visualized using a UV transilluminator. An example of gel electrophoresis results can be visualized in Figure 1.
The accuracy of the method was demonstrated by the lack of contamination (no amplicons in the negative control band) and the presence of positive control bands corresponding to all six DNA fragments of known molecular weight included in the marker used to determine the size of the amplification products. Also, internal controls indicated proper PCR amplification in each microtube (see Figure 1).
About 74 (33.03%) of the 224 subjects, both symptomatic and asymptomatic, were positive for one or more STDs. All six pathogen agents were detected with the multiplex PCR kit, several double or triple infections demonstrating associative patterns. The following double associations were detected in the study:
The bacteria most frequently identified in the analyzed urine samples was
Some studies recommended that all sexually active women should be tested, pregnant ones or those that have experienced miscarriages in particular, given the high prevalence of
A recent study in Korea  found a 3.3% infection rate for
In a study conducted in the United States,
5. Advantages of the multiplex PCR method
The advantages of the multiplex PCR method were assessed by various researchers comparing the diagnosis results obtained with different other conventional methods. The sensitivity and specificity of these methods were analyzed on different specimens in comparison to traditional methods, some of these considered to be the gold standard in diagnosis [36, 43, 46, 51]. Although this method can sometimes increase the workload, implying more protocol steps (DNA extraction, purification—if necessary, PCR amplification, electrophoresis), it still holds a huge advantage in terms of the exact diagnosis of STD. Except for special circumstances, such as when testing susceptibility to antibiotics of the various bacterial agents, NATTs have become the most sought after tests in STD detection. Regarding bacterial resistance to antibiotics, the DNA extracted and identified as belonging to such bacteria may be genetically tested to determine the genes that induce resistance to antibiotics or the different mutations that cause such resistance.
Although the symptoms are often similar, the treatment may differ depending on the STD agent detected. Without an accurate diagnosis, treatment is often ineffective. Clinicians often employ point‐of‐care (POC) tests to diagnose vaginal infections and STDs. For example, most clinicians rely on Amsel's clinical criteria in defining bacterial vaginosis: increased vaginal pH, the presence of amines, clue cells observed on wet mounts and homogeneous vaginal secretions . Although bacterial cultures and other methods are still widely used in STD detection, PCR tests are relevant, reproducible, sensitive and specific enough tools implying low costs and simplicity. In addition, the availability of commercial NAATs that include several STD pathogens allows more detailed studies regarding the relationship between such organisms in the etiology of these diseases. As well documented, all STDs occur in populations at high risk for other sexually transmitted infections. Their presence in a given individual is a marker for high‐risk behaviors and coincidental infections should be sought after, as well as other more serious sexually transmitted diseases such as HIV.
An advantage of this method is its great sensitivity consisting in the detection of STD pathogens in individuals who have previously been tested with conventional methods and came out with negative results.
The greatest advantage of this method is the detection of double or multiple coinfections. This is an important argument for promoting multiplex testing in the same sample. These results could have important implications in epidemiology and treatment by improvements in the accuracy of determining the possible synergies and interactions between such microorganisms.
STD screening for certain categories of population (e.g., patients attending sexual health clinics, infertile persons or women who had miscarriages) is necessary not only to identify symptomatic persons in order to diagnose and treat their infection but also to identify asymptomatic individuals who serve as possible infection carriers in order to reduce morbidity and help controlling these STDs. The multiplex PCR method provides a good opportunity to argue for STD screening.
Holmes K.K., Handsfield H.H. Sexually transmitted diseases: overview and clinical approach. In: Harrison’s Principles of Internal Medicine. 14th ed. Bucuresti: Teora; 2003. p. 881–893.
Salah R.M., Allam A.M., Magdy A.M., Mohamed A.S. Bacterial vaginosis and infertility: cause or association?. Eur J Obstet Gynecol Reprod Biol. 2013; 167(1):59–63. doi: 10.1016/j.ejogrb.2012.10.031
Korenromp E.L., White R.G., Orroth K.K., Bakker R., Kamali A., Serwadda D., et al. Determinants of the impact of sexually transmitted infection treatment on prevention of HIV infection: a synthesis of evidence from the Mwanza, Rakai, and Masaka intervention trials. J Infect Dis. 2005; 191(1):168–178. doi: 10.1086/425274
ECDC.Annual epidemiological report on communicable diseases in Europe. Stockholm: European Centre for Disease Prevention and Control. 2010
McGregor J.A., French J.I. Chlamydia trachomatisinfection during pregnancy. Am J Obstet Gynecol. 1991; 164(6):1782–1789. doi:10.1016/0002‐9378(91)90560‐E
Witkin S.S., Linhares I.M. Chlamydia trachomatisin subfertile women undergoing uterine instrumentation. Hum Reprod. 2002; 17(8):1938–1941
Hafner L.M., McNeilly C. Vaccines for Chlamydia infections of the female genital tract. Future Microbiol. 2008; 3(1):67–77. doi:10.2217/17460922.214.171.124
Anaes. Evaluation of screening low urogenital Chlamydia trachomatisin France. Paris: National Accreditation and Health Evaluation Agency. 2003
World Health Organization (WHO). Prevalence and incidence in 2005 of selected sexually transmitted infections. In: Chlamydia trachomatis, Neisseria gonorrhoeae, syphilis and Trichomonas vaginalis: methods and results used by WHO to generate 2005 estimates. WHO, Geneva. 2011
Li S.Y. Global transmission of multiple‐drug resistant Neisseria gonorrhoeaestrains refractive to cephalosporin treatment. J Formos Med Assoc. 2012; 111(9):463–464. doi:10.1016/j.jfma.2012.03.004
Querci M., Rombini F., Spinola L., Boutureira M., Yetman M., Reyes D., et al. Neisseria gonorrhoeaeinfective endocarditis. A case report. Int J Antimicrob Ag 41S1. 2013; S1–S34:P53.
Toyer A.L., Trignol‐Viguier N., Mereghetti L., Joly B., Blin E., Body G., et al. Interest of simultaneous Chlamydia trachomatisand Neisseria gonorrhoeaescreening at the time of preabortion consultation. Contraception. 2012; 86:572–576. doi:10.1016/j.contraception.2012.04.012
Gilbert R.O., Elia G., Beach D.H., Klaessig S., Singh B.N. Cytopathogenic effect of Trichomonas vaginalison human vaginal epithelial cells cultured in vitro. Infect Immun. 2000; 68(7):4200–4206.
Cudmore S.L., Garber G.E. Prevention or treatment: the benefits of Trichomonas vaginalisvaccine. J Infect Public Health. 2010; 3(2):47–53. doi:10.1016/j.jiph.2010.01.003
Cotch M.F., Pastorek J.G., Nugent R.P., Hillier S.L., Gibbs R.S., Martin D.H., et al. Trichomonas vaginalisassociated with low birth weight and preterm delivery. The Vaginal Infections and Prematurity Study Group. Sex Transm Dis. 1997; 24(6):353–360.
Noël J.C., Fayt I., Munoz M.R.R., Simon P., Engohan‐Aloghe C. High prevalence of high‐risk human papillomavirus infection among women with Trichomonas vaginalisinfection on monolayer cytology. Arch Gynecol Obstet. 2010; 282(5):503–505. doi:10.1007/s00404‐009‐1291‐x
Mayer K.H., Bush T., Henry K., Overton E.T., Hammer J., Richardson J., et al. Ongoing sexually transmitted disease acquisition and risk‐taking behavior among US HIV‐infected patients in primary care: implications for prevention interventions. Sex Transm Dis. 2012; 39(1):1–7. doi:10.1097/OLQ.0b013e31823b1922
Seña A.C., Miller W.C., Hobbs M.M., Schwebke J.R., Leone P.A., Swygard H., et al. Trichomonas vaginalisinfection in male sexual partners: implications for diagnosis, treatment, and prevention. Clin Infect Dis. 2007; 44(1):13–22. doi: 10.1086/511144
Pal C., Bandyopadhyay U. Redox‐active antiparasitic drugs. Antioxid Redox Signal. 2012; 17(4):555–582. doi:10.1089/ars.2011.4436
Sobel J.D., Nagappan V., Nyirjesy P. Metronidazole‐resistant vaginal trichomoniasis—an emerging problem. N Engl J Med. 1999; 341(4):292–293. doi:10.1056/NEJM199907223410417
Baka S., Kouskouni E., Antonopoulou S., Sioutis D., Papakonstantinou M., Hassiakos D., et al. Prevalence of Ureaplasma urealyticumand Mycoplasma hominisin women with chronic urinary symptoms. Urology. 2009; 74(1):62–66. doi: 10.1016/j.urology.2009.02.014
Wetmore C.M., Manhart L.E., Lowens M.S., Golden M.R., Whittington W.L., Xet‐Mull A.M., et al. Demographic, behavioral, and clinical characteristics of men with nongonococcal urethritis differ by etiology: a case‐comparison study. Sex Transm Dis. 2011; 38(3):180–186. doi:10.1097/OLQ.0b013e3182040de9
Manhart L.E., Holmes K.K., Hughes J.P., Houston L.S., Totten P.A. Mycoplasma genitaliumamong young adults in the United States: an emerging sexually transmitted infection. Am J Public Health. 2007; 97(6):1118–1125. doi: 10.2105/AJPH.2005.074062
Gupta A., Gupta A., Gupta S., Mittal A., Chandra P., Gill A.K. Correlation of mycoplasma with unexplained infertility. Arch Gynecol Obstet. 2009; 280(6):981–985. doi: 10.1007/s00404‐009‐1042‐z
Taylor‐Robinson D. The role of mycoplasmas in pregnancy outcome. Best Pract Res Clin Obstet Gynaecol. 2007; 21(3):425–438. doi:10.1016/j.bpobgyn.2007.01.011
Reichart M., Kahane I., Bartoov B. In vivo and in vitro impairment of human and ram sperm nuclear chromatin integrity by sexually transmitted Ureaplasma urealyticuminfection. Biol Reprod. 2000; 63(4):1041–1048.
Lee J.S., Kim K.T., Lee H.S., Yang K.M., Seo J.T., Choe J.H. Concordance of Ureaplasma urealyticumand Mycoplasma hominisin infertile couples: impact on semen parameters. Urology. 2013; 81(6):1219–1224. doi: 10.1016/j.urology.2013.02.044
Mirdh P.A., Paavonen J., Puolakkainen M, editors. Chlamydia. New York: Plenum Press Publishing Co; 1989.
Bianchi A. Chlamydia trachomatis et Chlamydia pneumoniae: diagnostic problems. Med Mal Infect. 1999; 29(Suppl. 1):38–50.
Mol B.W., Dijkman B., Wertheim P., Lijmer J., van der Veen F., Bossuyt P.M. The accuracy of serum chlamydial antibodies in the diagnosis of tubal pathology: a meta‐analysis. Fertil Steril. 1997; 67(6):1031–1037.
Dhama K., Karthik K., Chakraborty S., Tiwari R., Kapoor S., Kumar A., et al. Loop‐mediated isothermal amplification of DNA (LAMP): a new diagnostic tool lights the world of diagnosis of animal and human pathogens: a review. Pak J Biol Sci. 2014; 17(2):151–166.
Van Dyck E., Ieven M., Pattyn S., Van Damme L., Laga M. Detection of Chlamydia trachomatisand Neisseria gonorrhoeaeby enzyme immunoassay, culture, and three nucleic acid amplification tests. J Clin Microbiol. 2001; 39(5):1751–1756. doi: 10.1128/JCM.39.5.1751‐1756.2001
Whiley D.M., Tapsall J.W., Sloots T.P. Nucleic acid amplification testing for Neisseria gonorrhoeae: an ongoing challenge. J Mol Diagn. 2006; 8(1):3–15. doi: 10.2353/jmoldx.2006.050045
Gould S.B., Woehle C., Kusdian G., Landan G., Tachezy J., Zimorski V., et al. Deep sequencing of Trichomonas vaginalisduring the early infection of vaginal epithelial cells and amoeboid transition. Int J Parasitol. 2013; 43(9):707–719. doi:10.1016/j.ijpara.2013.04.002
Kissinger P. Trichomonas vaginalis: a review of epidemiologic, clinical and treatment issues. BMC Infect Dis. 2015; 15(1):307. doi:10.1186/s12879‐015‐1055‐0
Diaz N., Dessì D., Dessole S., Fiori P.L., Rappelli P. Rapid detection of coinfections by Trichomonas vaginalis, Mycoplasma hominis, and Ureaplasma urealyticumby a new multiplex polymerase chain reaction. Diagn Microbiol Infect Dis. 2010; 67(1):30–36. doi:10.1016/j.diagmicrobio.2009.12.022
Gatski M., Mena L., Levison J., Clark R.A., Henderson H., Schmidt N., et al. Patient‐delivered partner treatment and Trichomonas vaginalisrepeat infection among human immunodeficiency virus‐infected women. Sex Transm Dis. 2010; 37(8):502–505. doi:10.1097/OLQ.0b013e3181d891fc
Hobbs M.M., Lapple D.M., Lawing L.F., Schwebke J.R., Cohen M.S., Swygard H., et al. Methods for detection of Trichomonas vaginalisin the male partners of infected women: implications for control of trichomoniasis. J Clin Microbiol. 2006; 44(11):3994–3999. doi:10.1128/JCM.00952‐06
Brooks A.E., Parsamand T., Kelly R.W., Simoes‐Barbosa A. An improved quantitative method to assess adhesive properties of Trichomonas vaginalisto host vaginal ectocervical cells using flow cytometry. J Microbiol Methods. 2013; 92(1):73–78. doi:10.1016/j.mimet.2012.10.011
de Waaij D.J., Ouburg S., Dubbink J.H., Peters R.P.H., Morre S.A. Evaluation of Prestoplus assay and LightMix kit Trichomonas vaginalisassay for detection of Trichomonas vaginalisin dry vaginal swabs. J Microbiol Meth. 2016; 127:102–104. doi:10.1016/j.mimet.2016.06.002
Waites K.B., Talkington D. New developments of human diseases due to mycoplasmas. In: Blanchard A., Browning G., editors. Mycoplasmas: molecular biology, pathogenicity and strategies for control. Horizon bioscience; 2005. p. 289–354.
Ross J.D., Jensen J.S. Mycoplasma genitaliumas a sexually transmitted infection: implications for screening, testing, and treatment. Sex Transm Infect. 2006; 82(4):269–271. doi:10.1136/sti.2005.017368
Samra Z., Rosenberg S., Madar‐Shapiro L. Direct simultaneous detection of 6 sexually transmitted pathogens from clinical specimens by multiplex polymerase chain reaction and auto‐capillary electrophoresis. Diagn Microbiol Infect Dis. 2011; 70(1):17–21. doi:10.1016/j.diagmicrobio.2010.12.001
Baseman J.B., Cagle M., Korte J.E., Herrera C., Rasmussen W.G., Baseman J.G., et al. Diagnostic assessment of Mycoplasma genitaliumin culture‐positive women. J Clin Microbiol. 2004; 42(1):203–211.
Barré A., de Daruvar A., Blanchard A. MolliGen, a database dedicated to the comparative genomics of Mollicutes. Nucleic Acids Res. 2004; 32:D307–D310. doi: 10.1093/nar/gkh114
Choe H.S., Lee D.S., Lee S.J., Hong S.H., Park D.C., Lee M.K., et al. Performance of Anyplex™ II multiplex real‐time PCR for the diagnosis of seven sexually transmitted infections: comparison with currently available methods. Int J Infect Dis. 2013; 17(12):1134–1140. doi:10.1016/j.ijid.2013.07.011
Citti C., Blanchard A. Mycoplasmas and their host: emerging and re‐emerging minimal pathogens. Trends Microbiol. 2013; 21(4):196–203. doi: 10.1016/j.tim.2013.01.003
O'Byrne P. Self‐directed sexually transmitted infection testing: providing noninvasive sexual health services. Appl Nurs Res. 2011; 24(1):17–21. doi: 10.1016/j.apnr.2009.02.005
Ostergaard L. Diagnosis of urogenital Chlamydia trachomatisinfection by use of DNA amplification. APMIS Suppl. 1999; 89:5–36.
Fang J., Husman C., DeSilva L., Chang R., Peralta L. Evaluation of self‐collected vaginal swab, first void urine, and endocervical swab specimens for the detection of Chlamydia trachomatisand Neisseria gonorrhoeaein adolescent females. J Pediatr Adolesc Gynecol. 2008; 21(6):355–360. doi: 10.1016/j.jpag.2008.03.010
Lee S.J., Park D.C., Lee D.S., Choe H.S., Cho Y.H. Evaluation of Seeplex® STD6 ACE detection kit for the diagnosis of six bacterial sexually transmitted infections. J Infect Chemother. 2012; 18(4):494–500. doi:10.1007/s10156‐011‐0362‐7
Sturm‐Ramirez K., Brumblay H., Diop K., Guèye‐Ndiaye A., Sankalé J.L., Thior I., et al. Molecular epidemiology of genital Chlamydia trachomatisinfection in high‐risk women in Senegal, West Africa. J Clin Microbiol. 2000; 38(1):138–145.
Krõlov K., Frolova J., Tudoran O., Suhorutsenko J., Lehto T., Sibul H., et al. Sensitive and rapid detection of Chlamydia trachomatisby recombinase polymerase amplification directly from urine samples. J Mol Diagn. 2014; 16(1):127–135. doi: 10.1016/j.jmoldx.2013.08.003
U.S. Preventive Services Task Force. Screening for chlamydial infection: U.S. Preventive Services Task Force recommendation statement. Ann Intern Med. 2007; 147(2):128–134.
U.S. Preventive Services Task Force. Screening for Gonorrhea: recommendation statement. Ann Fam Med. 2005; 3(3):263–267. doi:10.1370%2Fafm.337
Stary A., Stary G. Sexually transmitted infection. In: Callen J.P., Cerroni L., Heymann W.R., Hruza G., Mancini A.J., Patterson J.W., et al, editors. Dermatology. 3rd ed. Elsevier; 2012. p. 1379–1383.
Keane F.E., Thomas B.J., Gilroy C.B., Renton A., Taylor‐Robinson D. The association of Mycoplasma hominis, Ureaplasma urealyticumand Mycoplasma genitaliumwith bacterial vaginosis: observations on heterosexual women and their male partners. Int J STD AIDS. 2000; 11(6):356–360.
Kim S.R., Kim J.H., Gu N.Y., Kim Y.S., Hong Y.C., Ryu J.S. Prevalence of trichomoniasis by PCR in women attending health screening in Korea. Korean J Parasitol. 2016; 54(2):187–190. doi:10.3347/kjp.2016.54.2.187
Waites K.B., Katz B., Schelonka R.L. Mycoplasmas and ureaplasmas as neonatal pathogens. Clin Microbiol Rev. 2005; 18(4):757–789. doi:10.1128/CMR.18.4.757‐789.2005
Amsel R., Totten P.A., Spiegel C.A., Chen K.C., Eschenbach D., Holmes K.K. Nonspecific vaginitis. Diagnostic criteria and microbial and epidemiologic associations. Am J Med. 1983; 74(1):14–22.