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Variable lipoprotein haemagglutinin (vlhA) gene sequence typing of mainly Dutch Mycoplasma synoviae isolates: comparison with vlhA sequences from Genbank and with amplified fragment length polymorphism analysis

机译:荷兰滑膜支原体分离株的可变脂蛋白血凝素(vlhA)基因序列类型:与Genbank的vlhA序列比较以及扩增的片段长度多态性分析

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摘要

Molecular typing techniques with sufficient discriminatory power are required to better understand the transmission of Mycoplasma synoviae, a poultry pathogen with increasing clinical and economic relevance. A promising molecular technique is polymerase chain reaction and subsequent sequencing based on the conserved 5 ' region of the M. synoviae variable lipoprotein and haemagglutinin (vlhA) gene. This technique was used for genotyping 27 mainly Dutch M. synoviae isolates from different organs of various categories of poultry housed on different farms and collected during a period of 10 years. The obtained vlhA sequences were compared with those of M. synoviae strains from Genbank and data obtained by amplified fragment length polymorphism (AFLP). Grouping based on 100% similarity revealed nine genotypes. Some isolates had identical vlhA gene sequences although they originated from different geographical areas, different years and organs. AFLP analysis results largely confirmed the results obtained by vlhA sequence typing. Our findings raise concern regarding the discriminatory power of these techniques for its use in molecular epidemiology of Dutch M. synoviae isolates and for the differentiation between M. synoviae vaccine strains and field isolates, and indicate that molecular typing based on additional markers should be considered.
机译:为了更好地了解滑膜支原体的传播,需要使用具有足够辨别力的分子分型技术,该支原体是一种具有越来越高的临床和经济意义的家禽病原体。一种有前途的分子技术是聚合酶链反应和基于滑膜支原体可变脂蛋白和血凝素(vlhA)基因的保守5'区的后续测序。该技术用于对来自不同农场的不同类别家禽的不同器官的27种主要荷兰滑膜支原体分离株进行基因分型,并在10年内进行了收集。将获得的vlhA序列与Genbank的滑膜支原体菌株进行比较,并通过扩增的片段长度多态性(AFLP)获得数据。基于100%相似性的分组揭示了9个基因型。尽管某些分离株来自不同的地理区域,不同的年份和器官,但它们具有相同的vlhA基因序列。 AFLP分析结果在很大程度上证实了通过vlhA序列分型获得的结果。我们的发现引起关注这些技术在荷兰滑膜分支杆菌分离株的分子流行病学中的应用以及在滑膜分支杆菌疫苗菌株和田间分离株之间的区分方面的歧视性,并表明应考虑基于其他标记的分子分型。

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