title: Simultaneous detection of different respiratory virus by a multiplex reverse transcription polymerase chain reaction combined with flow-through reverse dot blotting assay.
abstract: Cell culture and immunofluorescence (IF) assays have been traditionally used for the laboratory diagnosis of respiratory viral infections, but these assays have a low sensitivity and are time consuming. We developed a multiplex reverse transcription polymerase chain reaction combined with flow-through reverse dot blotting (mRT-PCR-FT-RDB) assay for the simultaneous detection of influenza virus type A including H5 subtype and H9 subtype, influenza virus type B, parainfluenza virus types 1 and 3, respiratory syncytial virus, human rhinovirus, and human coxsackievirus. In comparison with viral culture and IF assay as the gold standard method, the mRT-PCR-FT-RDB assay gave a sensitivity and a specificity of 100% and 98%. The high sensitivity and specificity, the rapid result turnaround time, and the reduced expense of the mRT-PCR-FT-RDB assay compared with viral culture and IF assay suggest that this assay would be a significant improvement over traditional ones for the detection of respiratory viruses in a clinical laboratory.
authors.affiliation: Department of Cell Biology and Genetics, School of Basic Science, Guangzhou Medical College, Guangzhou, Guangdong, China.
authors.forename: Pei-qiong
authors.initials: PQ
authors.lastname: Li
authors.forename: Zi-feng
authors.initials: ZF
authors.lastname: Yang
authors.forename: Jing-xian
authors.initials: JX
authors.lastname: Chen
authors.forename: Claude P
authors.initials: CP
authors.lastname: Muller
authors.forename: Jun
authors.initials: J
authors.lastname: Zhang
authors.forename: Dan-fen
authors.initials: DF
authors.lastname: Wang
authors.forename: Ren
authors.initials: R
authors.lastname: Zhang
authors.forename: Yun-shao
authors.initials: YS
authors.lastname: He
mesh_terms: D000328:Adult; D000818:Animals; D002460:Cell Line; D005260:Female; D005455:Fluorescent Antibody Technique; D006801:Humans; D015151:Immunoblotting / Q000379:methods*; D008297:Male; D008875:Middle Aged; D009305:Nasopharynx / Q000821:virology; D012367:RNA, Viral / Q000032:analysis / Q000302:isolation & purification; D012141:Respiratory Tract Infections / Q000175:diagnosis* / Q000821:virology*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013997:Time Factors; D014776:Virus Cultivation; D014777:Virus Diseases / Q000175:diagnosis / Q000821:virology; D014780:Viruses / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D012367:RNA, Viral