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Memórias do Instituto Oswaldo Cruz
Fundação Oswaldo Cruz, Fiocruz
ISSN: 1678-8060
EISSN: 1678-8060
Vol. 92, No. 3, 1997, pp. 389-393
Bioline Code: oc97071
Full paper language: English
Document type: Research Article
Document available free of charge

Memórias do Instituto Oswaldo Cruz, Vol. 92, No. 3, 1997, pp. 389-393

 en Analysis of respiratory syncytial virus in clinical samples by reverse transcriptase-polymerase chain reaction restriction mapping
Angel Valdivia; Clara Savon; Danay Chacon; Luis Sarmiento; Luis Morier; Anselmo Otero; Yudira Soto; Suset Oropesa & Angel Goyenechea

Abstract

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.

Keywords
respiratory syncytial virus - polymerase chain reaction - restriction mapping - diagnosis

 
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