Analysis of respiratory syncytial virus in clinical samples by reverse transcriptase-polymerase chain reaction restriction mapping.
The aim of this study was to develop a polymerase chain reaction (PCR) for the detection of respiratory syncytial virus (RSV) genomes. The primers were designed from published sequences and selected from conserved regions of the genome encoding for the N protein of subgroups A and B of RSV. PCR was applied to 20 specimens from children admitted to the respiratory ward of "William Soler" Pediatric Hospital in Havana City with a clinical diagnosis of bronchiolitis. The PCR was compared with viral isolation and with an indirect immunofluorescence technique that employs monoclonal antibodies of subgroups A and B. Of 20 nasopharyngeal exudates, 10 were found positive by the three assayed methods. In only two cases, samples that yielded positive RNA-PCR were found negative by indirect immunofluorescence and cell culture. Considering viral isolation as the "gold standard" technique, RNA-PCR had 100% sensitivity and 80% specificity. RNA-PCR is a specific and sensitive technique for the detection of the RSV genome. Technical advantages are discussed.
Valdivia
A
A
Instituto de Medicina Tropical Pedro Kouri, La Habana, Cuba.
Savón
C
C
Chacón
D
D
Sarmiento
L
L
Morier
L
L
Otero
A
A
Soto
Y
Y
Oropesa
S
S
Goyenechea
A
A
D006801:Humans; D016133:Polymerase Chain Reaction; D012194:RNA-Directed DNA Polymerase; D018357:Respiratory Syncytial Virus Infections / Q000175:diagnosis*; D018113:Respiratory Syncytial Virus, Human / Q000235:genetics*; D015183:Restriction Mapping
D012194:RNA-Directed DNA Polymerase