Chapters authored
Multiplex Polymerase Chain Reaction Assay for Early Diagnosis of Viral Infection
Viral reactivation is one of the most serious complications for immunocompromised patients. Under immunosuppressive conditions, some viruses can be reactivated solely or simultaneously and may thus cause life-threatening infection. Therefore, the prompt and proper diagnosis of viral reactivation is important for the initiation of preemptive therapy. For this purpose, we recently developed a multiplex-virus polymerase chain reaction (PCR) assay. The multiplex PCR assay is designed to qualitatively measure the genomic DNA of 12 viruses at once: cytomegalovirus (CMV), human herpesvirus type 6 (HHV-6), HHV-7, HHV-8, Epstein-Barr virus (EBV), varicella-zoster virus (VZV), BK virus (BKV), JC virus (JCV), parvovirus B19 (ParvoB19), herpes simplex virus type 1 (HSV-1), HSV-2, and hepatitis B virus (HBV). When a specific PCR signal is obtained, the viral load is determined by a quantitative real-time PCR. The qualitative multiplex and quantitative real-time PCR procedures take only 3 hours to complete. With this assay system, we can identify viremia at the early stage and thereby prevent it from progressing to overt and symptomatic viral infection in immunocompromised patients, such as those receiving hematopoietic stem cell transplantation.
Part of the book: Polymerase Chain Reaction for Biomedical Applications