Rapid detection of different RNA respiratory virus species by multiplex RT-PCR: application to clinical specimens.
BACKGROUND
The polymerase chain reaction (PCR) applied in diagnostic and epidemiologic investigations is very useful for sensitivity, specificity and time saving.


OBJECTIVE
We have developed a method for the detection of genomic RNA of two different species of virus, the influenza A virus (IA) and the respiratory syncytial virus (RS), which are responsible for clinical similarities. We applied this multiplex RT-PCR protocol on clinical specimens.


METHODS
We describe a method which allows rapid diagnosis by performing a single retro-transcriptase (RT) reaction associated with the PCR (multiplex RT-PCR) on different genomes in a single sample. We have evaluated the sensitivity and the specificity of the multiplex test on positive controls, then, on RNA extracted from clinical specimens harvested from 15 children with respiratory symptoms during the spring-winter season 1997.


CONCLUSIONS
The multiplex RT-PCR protocol, applied to respiratory specimens, allows the investigation of RNA IA virus and RS virus in a single sample at the same time. The detection of the etiologic viral agent is rapid and it is possible to evaluate incidental simultaneous infections.
Valassina
M
M
Department of Molecular Biology, University of Siena, Italy.
Cuppone
A M
AM
Cusi
M G
MG
Valensin
P E
PE
D000818:Animals; D002460:Cell Line; D002648:Child; D002675:Child, Preschool; D004285:Dogs; D006801:Humans; D007223:Infant; D009980:Influenza A virus / Q000235:genetics / Q000302:isolation & purification*; D007251:Influenza, Human / Q000175:diagnosis* / Q000473:pathology; D009305:Nasopharynx / Q000378:metabolism / Q000821:virology; D016133:Polymerase Chain Reaction / Q000379:methods*; D018357:Respiratory Syncytial Virus Infections / Q000175:diagnosis*; D018113:Respiratory Syncytial Virus, Human / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D013997:Time Factors; D014407:Tumor Cells, Cultured