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利用聚合酶链反应(PCR)结合限制性酶切分析对印度北部家禽腺病毒野外分离株进行特征分析。

The use of PCR combined with restriction enzyme analysis to characterize fowl adenovirus field isolates from northern India.

作者信息

Singh A, Oberoi M S, Grewal G S, Hafez H M, Hess M

机构信息

Department of Veterinary Pathology, College of Veterinary Science, Punjab Agricultural University, Ludhiana, India.

出版信息

Vet Res Commun. 2002 Oct;26(7):577-85. doi: 10.1023/a:1020299700907.

Abstract

Ten fowl adenoviruses (FAVs), isolated from suspected cases of inclusion body hepatitis (IBH) in quails and broilers, were characterized by a hexon-based polymerase chain reaction (PCR) combined with restriction enzyme analysis (REA) of the amplified DNA fragments. All the isolates could be detected using H1/H2 and H3/H4 primer sets. Amplification of DNA with H1/H2 and H3/H4 primer sets resulted in fragments of approximately 1219 bp and 1319 bp, respectively. HaeII digestion of the H1/ H2 PCR products and HpaII digestion of the H3/H4 PCR products characterized all the isolates in FAV groups, known from genomic typing using the whole DNA. For some of the isolates, neutralization tests were used to confirm these results. The results revealed that, as well as FAV serotype 1, which is the sole member of DNA group A, FAVs of DNA group E are also associated with IBH in poultry in northern India. The FAV specific PCR combined with REA was found to be very useful in investigating the epidemiological situation in the field. It was even possible to define mixed infections with more than one FAV.

摘要

从鹌鹑和肉鸡中疑似包涵体肝炎(IBH)病例分离出的10株禽腺病毒(FAV),通过基于六邻体的聚合酶链反应(PCR)结合对扩增DNA片段的限制性酶切分析(REA)进行了特征鉴定。使用H1/H2和H3/H4引物对均可检测到所有分离株。用H1/H2和H3/H4引物对扩增DNA分别得到约1219 bp和1319 bp的片段。对H1/H2 PCR产物进行HaeII酶切以及对H3/H4 PCR产物进行HpaII酶切,可对使用全基因组进行基因分型已知的FAV组中的所有分离株进行特征鉴定。对于部分分离株,使用中和试验来确认这些结果。结果显示,除了作为A组DNA唯一成员的1型FAV外,E组DNA的FAV也与印度北部家禽的IBH有关。发现FAV特异性PCR结合REA在调查该领域的流行病学情况时非常有用。甚至有可能确定由一种以上FAV引起的混合感染。

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