title: A novel real-time PCR system for simultaneous detection of human viruses in clinical samples from patients with uncertain diagnoses.
abstract: A novel simultaneous detection system for human viruses was developed using a real-time polymerase chain reaction (PCR) system to identify causes of infection in clinical samples from patients with uncertain diagnoses. This system, designated as the "multivirus real-time PCR," has the potential to detect 163 human viruses (47 DNA viruses and 116 RNA viruses) in a 96-well plate simultaneously. The specificity and sensitivity of each probe-primer set were confirmed with cells or tissues infected with specific viruses. The multivirus real-time PCR system showed profiles of virus infection in 20 autopsies of acquired immunodeficiency syndrome patients, and detected frequently TT virus, cytomegalovirus, human herpesvirus 6, and Epstein-Barr virus in various organs; however, RNA viruses were detected rarely except for human immunodeficiency virus-1. Pathology samples from 40 patients with uncertain diagnoses were examined, including cases of encephalitis, hepatitis, and myocarditis. Herpes simplex virus 1, human herpesvirus 6, and parechovirus 3 were identified as causes of diseases in four cases of encephalitis, while no viruses were identified in other cases as causing disease. This multivirus real-time PCR system can be useful for detecting virus in specimens from patients with uncertain diagnoses.
authors.affiliation: Department of Pathology, National Institute of Infectious Diseases, Shinjuku, Tokyo, Japan. katano@nih.gov.jp
authors.forename: Harutaka
authors.initials: H
authors.lastname: Katano
authors.forename: Motofumi
authors.initials: M
authors.lastname: Kano
authors.forename: Tomoyuki
authors.initials: T
authors.lastname: Nakamura
authors.forename: Takayuki
authors.initials: T
authors.lastname: Kanno
authors.forename: Hideki
authors.initials: H
authors.lastname: Asanuma
authors.forename: Tetsutaro
authors.initials: T
authors.lastname: Sata
mesh_terms: D000163:Acquired Immunodeficiency Syndrome / Q000175:diagnosis / Q000821:virology; D000328:Adult; D000368:Aged; D017931:DNA Primers; D004267:DNA Viruses / Q000235:genetics / Q000302:isolation & purification*; D004279:DNA, Viral / Q000032:analysis; D005260:Female; D015497:HIV-1 / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction / Q000379:methods*; D012328:RNA Viruses / Q000235:genetics / Q000302:isolation & purification*; D012367:RNA, Viral / Q000032:analysis; D012680:Sensitivity and Specificity; D014777:Virus Diseases / Q000175:diagnosis*; D014781:Viscera / Q000821:virology
chemical_list: D017931:DNA Primers; D004279:DNA, Viral; D012367:RNA, Viral