生物
病毒学
H5N1亚型流感病毒
实时聚合酶链反应
计算生物学
遗传学
基因
病毒
作者
Olga V. Petrauskene,M. A. Schumaker,Yvonne R. Thorstenson,Catherine Fearnley,T. Pavlidis,Sueh-Ning Liew,Jennifer Cork,Manohar R. Furtado,Philip R. Wakeley,Marek J. Slomka
出处
期刊:Avian Diseases
[BioOne (American Association of Avian Pathologists)]
日期:2010-03-01
卷期号:54 (s1): 686-689
被引量:4
标识
DOI:10.1637/8797-040109-resnote.1
摘要
New lyophilized real-time reverse transcription (RT)-PCR avian influenza detection assays were designed and tested. The M-gene assay detects all avian influenza virus (AIV) subtypes, and the H5 and H7 specific assays can discriminate the AIV subtypes H5 and H7 of Eurasian origin. The assays are formulated in a lyophilized bead format containing an internal positive control to monitor inhibitors in the reaction. Fifty-six AIV cultured isolates covering all 16 hemagglutinin types and 44 positive swabs from an outbreak of AIV in turkeys (H5N1 highly pathogenic avian influenza) were used to determine analytical performance and diagnostic sensitivity of these veterinary assays. The lyophilized real-time RT-PCR assays were demonstrated to be more sensitive than the wet assays, being able to detect down to 4 to 16 molecules of synthetic target RNA compared to 16 to 80 molecules for the corresponding wet assays. The diagnostic sensitivity of the lyophilized M-gene assay was determined to be 97.7% (43/44), whereas concurrent testing of these samples with the wet assay was only 86.3% sensitive (38/44). Using a panel of 19 noninfluenza respiratory and enteric pathogens, the analytical specificity of the M-gene assay was shown to be 100%. High diagnostic specificity of the assays was also confirmed by testing 496 negative swab samples from a combination of wild bird species and poultry.
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