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Molecular Variability of the Adhesin-Encoding Gene pvpA among Mycoplasma gallisepticum Strains and Its Application in Diagnosis

机译:鸡败血支原体菌株间粘附素编码基因pvpA的分子变异性及其在诊断中的应用

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摘要

Mycoplasma gallisepticum is an important pathogen of chickens and turkeys that causes considerable economic losses to the poultry industry worldwide. The reemergence of M. gallisepticum outbreaks among poultry, the increased use of live M. gallisepticum vaccines, and the detection of M. gallisepticum in game and free-flying song birds has strengthened the need for molecular diagnostic and strain differentiation tests. Molecular techniques, including restriction fragment length polymorphism of genomic DNA (RFLP) and PCR-based random amplification of polymorphic DNA (RAPD), have already been utilized as powerful tools to detect intraspecies variation. However, certain intrinsic drawbacks constrain the application of these methods. The main goal of this study was to determine the feasibility of using an M. gallisepticum-specific gene encoding a phase-variable putative adhesin protein (PvpA) as the target for molecular typing. This was accomplished using a pvpA PCR-RFLP assay. Size variations among PCR products and nucleotide divergence of the C-terminus-encoding region of the pvpA gene were the basis for strain differentiation. This method can be used for rapid differentiation of vaccine strains from field isolates by amplification directly from clinical samples without the need for isolation by culture. Moreover, molecular epidemiology of M. gallisepticum outbreaks can be performed using RFLP and/or sequence analysis of the pvpA gene.
机译:鸡支原体支原体是鸡和火鸡的重要病原体,对全世界的家禽业造成可观的经济损失。家禽中鸡毒支原体暴发的再次出现,活鸡毒支原体疫苗的使用增加以及在野味和自由飞翔的鸣禽中检出鸡毒支原体,这增加了对分子诊断和菌株鉴别测试的需求。分子技术,包括基因组DNA的限制性片段长度多态性(RFLP)和基于PCR的多态性DNA随机扩增(RAPD),已被用作检测种内变异的强大工具。但是,某些固有的缺点限制了这些方法的应用。这项研究的主要目标是确定使用编码相位可变推定粘附素蛋白(PvpA)的鸡毒支原体特异性基因作为分子分型目标的可行性。这是使用pvpA PCR-RFLP分析完成的。 PCR产物之间的大小变化和pvpA基因的C末端编码区的核苷酸差异是菌株分化的基础。通过直接从临床样品中扩增而无需通过培养分离,该方法可用于从野外分离株中快速区分疫苗株。而且,可以使用RFLP和/或pvpA基因的序列分析来进行鸡败血支原体暴发的分子流行病学。

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