Simultaneous detection of different respiratory virus by a multiplex reverse transcription polymerase chain reaction combined with flow-through reverse dot blotting assay.
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.
Li
Pei-qiong
PQ
Department of Cell Biology and Genetics, School of Basic Science, Guangzhou Medical College, Guangzhou, Guangdong, China.
Yang
Zi-feng
ZF
Chen
Jing-xian
JX
Muller
Claude P
CP
Zhang
Jun
J
Wang
Dan-fen
DF
Zhang
Ren
R
He
Yun-shao
YS
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*
D012367:RNA, Viral