title: Clinical Evaluation of a Single-Tube Multiple RT-PCR Assay for the Detection of 13 Common Virus Types/Subtypes Associated with Acute Respiratory Infection.
abstract: Respiratory viruses are among the most important causes of human morbidity and mortality worldwide, especially for infants and young children. In the past years, a few commercial multiplex RT-PCR assays have been used to detect respiratory viruses in spite of the high cost. In the present study, an improved single-tube multiplex reverse transcription PCR assay for simultaneous detection of 13 respiratory viruses was evaluated and compared with a previously reported two-tube assay as the reference method using clinical nasopharyngeal aspirates samples. Of 310 prospectively tested respiratory specimens selected from children hospitalized with acute respiratory illness, 226 (72.90%, 226/310) and 214 (69.03%, 214/310) positive for one or more viruses were identified by the single-tube and the two-tube assays, respectively, with combined test results showing good concordance (Kappa value = 0.874). Individually, the single-tube assay for adenovirus (Adv), human metapneumovirus (HMPV), human rhinovirus (HRV), parainfluenza virus type 1 (PIV1), parainfluenza virus type 3 (PIV3) and parainfluenza virus type 4 (PIV4) showed the significantly superior sensitivities to those of the two-tube assay. No false positives were found. In conclusion, our results demonstrates the one-tube assay revealed significant improvements over the two-tube assay in terms of the better sensitivity, more accurate quality control, less nonspecific amplification, more cost-effective and shorter turn-around time and will be a valuable tool for routine surveillance of respiratory virus infection in China.
authors.affiliation: Department of Pathophysiology, Guangzhou Medical University, Guangzhou city, Guangdong, China.|Key Laboratory for Medical Virology, National Health and Family Planning Commission, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Changping district, Beijing, China.
authors.forename: Dan
authors.initials: D
authors.lastname: Zhang
authors.affiliation: Pediatric Research Institute, Children's Hospital of Hebei Province, Hebei Medical University, Shijiazhuang, China.
authors.forename: Zhishan
authors.initials: Z
authors.lastname: Feng
authors.affiliation: Key Laboratory for Medical Virology, National Health and Family Planning Commission, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Changping district, Beijing, China.|Pediatric Research Institute, Children's Hospital of Hebei Province, Hebei Medical University, Shijiazhuang, China.
authors.forename: Mengchuan
authors.initials: M
authors.lastname: Zhao
authors.affiliation: Key Laboratory for Medical Virology, National Health and Family Planning Commission, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Changping district, Beijing, China.|Department of Infectious Diseases, Institute of Biomedicine, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.
authors.forename: Hao
authors.initials: H
authors.lastname: Wang
authors.affiliation: Pediatric Research Institute, Children's Hospital of Hebei Province, Hebei Medical University, Shijiazhuang, China.
authors.forename: Le
authors.initials: L
authors.lastname: Wang
authors.affiliation: Pediatric Research Institute, Children's Hospital of Hebei Province, Hebei Medical University, Shijiazhuang, China.
authors.forename: Shuo
authors.initials: S
authors.lastname: Yang
authors.affiliation: Pediatric Research Institute, Children's Hospital of Hebei Province, Hebei Medical University, Shijiazhuang, China.
authors.forename: Guixia
authors.initials: G
authors.lastname: Li
authors.affiliation: Department of Pathophysiology, Guangzhou Medical University, Guangzhou city, Guangdong, China.
authors.forename: Li
authors.initials: L
authors.lastname: Lu
authors.affiliation: Key Laboratory for Medical Virology, National Health and Family Planning Commission, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Changping district, Beijing, China.
authors.forename: Xuejun
authors.initials: X
authors.lastname: Ma
mesh_terms: D000208:Acute Disease; D000293:Adolescent; D002648:Child; D002681:China; D005260:Female; D006801:Humans; D007223:Infant; D008297:Male; D012141:Respiratory Tract Infections / Q000175:diagnosis / Q000235:genetics / Q000821:virology; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D014777:Virus Diseases / Q000175:diagnosis / Q000235:genetics; D014780:Viruses / Q000235:genetics*