Detection of human rhinovirus RNA in nasal washings by PCR
Identifieur interne : 001B19 ( Main/Exploration ); précédent : 001B18; suivant : 001B20Detection of human rhinovirus RNA in nasal washings by PCR
Auteurs : E. Arruda [États-Unis] ; F. G. Hayden [États-Unis]Source :
- Molecular and Cellular Probes [ 0890-8508 ] ; 1993.
English descriptors
Abstract
Abstract: A PCR assay was developed to detect human rhinovirus (HRV) RNA in nasal washings from individuals experimentally infected with HRV-39 or HRV strain Hanks. Total RNA was purified from samples stored in the presence of vanadyl ribonucleoside complex (VRC) by one of two methods: proteinase K digestion followed by multiple extractions with phenol/chloroform (PK-PC); or denaturation with guanidinium thiocyanate followed by one phenol/chloroform extraction (GTC). The limit of detection of HRV in nasal washings spiked with HRV-39 was lower with the GTC method (1 TCID50) than with the PK-PC method. In a study of 31 washings extracted by the PK-PC method, the sensitivity (93%) and negative predictive value (94%) were sub-optimal in comprison to cell culture. In a study of 60 nasal washings extracted by the GTC method, the number of samples positive by PCR (25) exceeded by two the number positive by isolation in cell culture. A GTC-based method for HRV RNA extraction in nasal washings was superior to a proteinase K-phenol/chloroform-based method in regard to sensitivity, consumption of reagents, material and time.
Url:
DOI: 10.1006/mcpr.1993.1055
Affiliations:
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