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A multiplex RT-PCR for detection of type A influenza virus and differentiation of avian H5, H7, and H9 hemagglutinin subtypes.

Abstract：

:A multiplex reverse transcriptase-polymerase chain reaction (mRT-PCR) was developed and optimized for the detection of type A influenza virus; the assay simultaneously differentiates avian H5, H7 and H9 hemagglutinin subtypes. Four sets of specific oligonucleotide primers were used in this test for type A influenza virus, H5, H7 and H9 heamagglutinin subtypes. The mRT-PCR DNA products were visualized by gel electrophoresis and consisted of fragments of 860 bp for H5, 634 bp for H7, 488 bp for H9 hemagglutinin subtypes, and 244 bp for type A influenza virus. The common set primers for type A influenza virus were able to amplify a 244 bp DNA band for any of the other subtypes of AIV. The mRT-PCR assay developed in this study was found to be sensitive and specific. Detection limit for PCR-amplified DNA products was 100 pg for the subtypes H5, H7, and H9 and 10 pg for type A influenza virus in all subtypes. No specific amplification bands of the same sizes (860, 634 and 488 bp) could be amplified for RNA of other influenza hemagglutinin subtypes, nor specific amplification bands of type A influenza (244 bp) for other viral or bacterial pathogens.

journal_name

Mol Cell Probes

journal_title

Molecular and cellular probes

authors

Xie Z,Pang YS,Liu J,Deng X,Tang X,Sun J,Khan MI

doi

10.1016/j.mcp.2006.01.003

subject

Has Abstract

pub_date

2006-06-01 00:00:00

pages

245-9

issue

3-4

eissn

0890-8508

issn

1096-1194

pii

S0890-8508(06)00016-8

journal_volume

20

pub_type

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