Heminested PCR Assay for Detection of Six Genotypes of Rabies and Rabies-related Viruses

Overview

Journal J Clin Microbiol

Specialty Microbiology

Date 1997 Nov 14

PMID 9350729

Citations 94

Authors

P R Heaton

P Johnstone

L M McElhinney

R Cowley

E OSullivan

J E Whitby

Affiliations

Soon will be listed here.

Abstract

A heminested reverse transcriptase PCR (hnRT-PCR) protocol which is rapid and sensitive for the detection of rabies virus and rabies-related viruses is described. Sixty isolates from six of the seven genotypes of rabies and rabies-related viruses were screened successfully by hnRT-PCR and Southern blot hybridization. Of the 60 isolates, 93% (56 of 60) were positive by external PCR, while all isolates were detected by heminested PCR and Southern blot hybridization. We also report on a comparison of the sensitivity of the standard fluorescent-antibody test (FAT) for rabies antigen and that of hnRT-PCR for rabies viral RNA with degraded tissue infected with a genotype 1 virus. Results indicated that FAT failed to detect viral antigen in brain tissue that was incubated at 37 degrees C for greater than 72 h, while hnRT-PCR detected viral RNA in brain tissue that was incubated at 37 degrees C for 360 h.

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