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应用随机引物聚合酶链反应对胸膜肺炎放线杆菌进行血清型鉴定

Serotype identification of Actinobacillus pleuropneumoniae by arbitrarily primed polymerase chain reaction.

作者信息

Hennessy K J, Iandolo J J, Fenwick B W

机构信息

Department of Pathology and Microbiology, College of Veterinary Medicine, Kansas State University, Manhattan 66506.

出版信息

J Clin Microbiol. 1993 May;31(5):1155-9. doi: 10.1128/jcm.31.5.1155-1159.1993.

Abstract

Rapid and accurate determination of the Actinobacillus pleuropneumoniae serotype involved in a disease outbreak is important both in limiting the severity of an outbreak and for tracing the source of the infecting organism. This study describes the use of arbitrarily primed polymerase chain reaction (AP-PCR) as a rapid, precise, and genetically based procedure to identify A. pleuropneumoniae. AP-PCR amplification of bacterial genomic DNA results in specific DNA profiles, which can be used to differentiate currently recognized serotypes. This technique is especially useful for identifying previously nontypeable and serologically cross-reactive A. pleuropneumoniae field isolates. Consecutive passages of isolates on different media, freezing, and subsequent infection of pigs did not alter the AP-PCR genomic profile. We propose the use of M13 and T3-T7 oligodeoxynucleotide primers for diagnostic and epidemiological identification of A. pleuropneumoniae by AP-PCR techniques.

摘要

快速准确地确定疾病暴发中涉及的胸膜肺炎放线杆菌血清型,对于限制疫情的严重程度以及追踪感染源都很重要。本研究描述了使用任意引物聚合酶链反应(AP-PCR)作为一种快速、精确且基于基因的方法来鉴定胸膜肺炎放线杆菌。细菌基因组DNA的AP-PCR扩增产生特定的DNA图谱,可用于区分目前公认的血清型。该技术对于鉴定先前无法分型和血清学交叉反应的胸膜肺炎放线杆菌田间分离株特别有用。分离株在不同培养基上连续传代、冷冻以及随后感染猪,均未改变AP-PCR基因组图谱。我们建议使用M13和T3-T7寡脱氧核苷酸引物,通过AP-PCR技术对胸膜肺炎放线杆菌进行诊断和流行病学鉴定。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/30c6/262895/c2fe38c18460/jcm00017-0152-a.jpg

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