利用全细胞、外膜和聚合酶链反应分型鉴定兔多杀性巴氏杆菌分离株。

Laboratory animal science Pub Date : 1999-10-01
S M Dabo, A W Confer, M Montelongo, Y S Lu
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引用次数: 0

摘要

目的:采用血清分型、十二烷基硫酸钠-聚丙烯酰胺凝胶电泳全细胞蛋白(WCPs)和外膜蛋白(OMPs)以及聚合酶链反应(PCR)指纹图谱对家兔多杀性巴氏杆菌分离物进行鉴定。方法:从ATCC(1)、Oklahoma(4)、Michigan(9)、Minnesota(7)和Texas(29) 5个来源分离50株。PCR指纹图谱采用两个微卫星探针(GTG)5和(GACA)4对M13和修改后的M13核心序列进行分析。结果:A血清组分离45株,d血清组分离5株。共有10株WCP型(w1 ~ w10)和10株OMP型(om1 ~ om10)变异(W1a)。引物M13噬菌体、修饰的M13噬菌体、(GTG)5和(GACA)4分别产生7、9、5和9种指纹类型。综合WCP、OMP和PCR指纹图谱结果得到39组,鉴别指数为0.98。PCR指纹图谱结果表明,不同地理位置的分离株间存在克隆相关性,但德克萨斯州分离株的PCR指纹图谱类型差异较大。结论:单引物PCR指纹图谱为家兔多杀假单胞菌分型提供了一种简便、快速的方法。常规分型和分子分型的结合增强了从巴氏杆菌病家兔分离的多杀假单胞菌的分化。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Characterization of rabbit Pasteurella multocida isolates by use of whole-cell, outer-membrane, and polymerase chain reaction typing.

Purpose: To characterize Pasteurella multocida isolates from laboratory rabbits using serotyping, sodium dodecyl sulfate-polyacrylamide gel electrophoresis of whole-cell proteins (WCPs) and outer-membrane proteins (OMPs), and polymerase chain reaction (PCR) fingerprinting.

Methods: Fifty isolates were obtained from five sources: ATCC (1), Oklahoma (4), Michigan (9), Minnesota (7), and Texas (29). The PCR fingerprinting was conducted using two minisatellite probes for M13 and a modified M13 core sequence and two microsatellite probes--(GTG)5 and (GACA)4.

Results: Forty-five isolates were serogroup A, and five were serogroup D. Ten WCP patterns (W1-W10) with one variation (W1a) and 10 OMP (OM1-OM10) patterns were found. Primers M13 phage, modified M13 phage, (GTG)5, and (GACA)4 generated 7, 9, 5, and 9 fingerprint types, respectively. Combination of WCP, OMP, and PCR fingerprint results yielded 39 groups with a discrimination index of 0.98. The PCR fingerprint results generally indicated clonal association among isolates within geographic locations except for the isolates from Texas, which varied markedly in PCR fingerprint types.

Conclusion: Single primer PCR fingerprinting provided a simple and rapid means of typing P. multocida isolates from laboratory rabbits. Combinations of conventional and molecular typing enhanced differentiation among P. multocida isolated from rabbits with pasteurellosis.

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