MULTIPLEX PCR DETECTION AND DIFFERENTIATION OF INFLUENZA A AND B VIRUSES IN CLINICAL SPECIMENS

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Authors
  1. Fisher, C.
  2. Evans, D.H.
Corporate Authors
Defence Research Establishment Suffield, Ralston ALTA (CAN);Guelph Univ, Guelph ONT (CAN) Dept of Molecular Biology and Genetics
Abstract
Influenza A and B viruses are multi-segmented RNA viruses responsible for sporadic epidemics of human influenza. They are currently diagnosed in respiratory secretions either by direct immunofluorescence (DIF) and by DIF coupled with cell culture techniques. In this study we examined whether nested PCR and reverse transcriptase reactions offers a viable alternative to detecting and typing these viruses. A collection of clinical specimens were obtained from the Alberta Laboratory of Public Health including influenza. A and B isolates plus isolates identified as containing respiratory syncytial virus, adenovirus, or parainfluenza virus. Culture negative specimens were used as negative controls. Two sets of nested PCR primers were found to efficiently detect A and B viruses in high-titre viral stocks and showed essentially no cross-reaction with samples known to contain heterologous viral species. Our ability to control contamination was shown by the failure to detect viral nucleic acids in specimens previously shown to lack any cultivable virus. The influenza A primer set designated AMP-1 and the influenza B primer set BMP-3 were also tested under multiplex conditions and still found to specifically amplify only the homologous virus. Using this simplified multiplex assay we were able to detect virus in 65% (13/20) of the DIF-positive influenza A isolates and 35% (7/20) of the DIF-positive influenza B specimens. TRUNCATED.
Keywords
Influenza A;Influenza B;Orthomyxoviridae;Viral detection;Polymerase chain reaction (PCR);Immunofluorescence;Viral sequences;Nucleic acid sequencing
Report Number
DRES-CR-95-10 — Contract Report
Date of publication
01 Feb 1995
Number of Pages
46
DSTKIM No
95-03268
CANDIS No
151745
Format(s):
Document Image stored on Optical Disk;Hardcopy

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