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应用随机扩增多态性 DNA 和蛋白图谱对副猪嗜血杆菌参考株和分离株的比较。

Comparison of Haemophilus parasuis reference strains and field isolates by using random amplified polymorphic DNA and protein profiles.

机构信息

Ruminant Diseases and Immunology, National Animal Disease Center, Agricultural Research Service, US Department of Agriculture, Ames, IA 50010, USA.

出版信息

BMC Microbiol. 2012 Jun 15;12:108. doi: 10.1186/1471-2180-12-108.

Abstract

BACKGROUND

Haemophilus parasuis is the causative agent of Glässer's disease and is a pathogen of swine in high-health status herds. Reports on serotyping of field strains from outbreaks describe that approximately 30% of them are nontypeable and therefore cannot be traced. Molecular typing methods have been used as alternatives to serotyping. This study was done to compare random amplified polymorphic DNA (RAPD) profiles and whole cell protein (WCP) lysate profiles as methods for distinguishing H. parasuis reference strains and field isolates.

RESULTS

The DNA and WCP lysate profiles of 15 reference strains and 31 field isolates of H. parasuis were analyzed using the Dice and neighbor joining algorithms. The results revealed unique and reproducible DNA and protein profiles among the reference strains and field isolates studied. Simpson's index of diversity showed significant discrimination between isolates when three 10 mer primers were combined for the RAPD method and also when both the RAPD and WCP lysate typing methods were combined.

CONCLUSIONS

The RAPD profiles seen among the reference strains and field isolates did not appear to change over time which may reflect a lack of DNA mutations in the genes of the samples. The recent field isolates had different WCP lysate profiles than the reference strains, possibly because the number of passages of the type strains may affect their protein expression.

摘要

背景

副猪嗜血杆菌是格拉泽氏病的病原体,也是健康猪群中的猪病原体。关于暴发疫情中野外菌株的血清分型报告描述,大约 30%的菌株是不可分型的,因此无法追踪。分子分型方法已被用作血清分型的替代方法。本研究旨在比较随机扩增多态性 DNA(RAPD)图谱和全细胞蛋白(WCP)裂解物图谱,作为区分副猪嗜血杆菌参考株和野外分离株的方法。

结果

使用 Dice 和邻接法分析了 15 株参考株和 31 株副猪嗜血杆菌野外分离株的 DNA 和 WCP 裂解物图谱。结果表明,在所研究的参考株和野外分离株中,DNA 和蛋白质图谱具有独特性和可重复性。当使用三种 10 碱基引物进行 RAPD 方法,以及同时使用 RAPD 和 WCP 裂解物分型方法时,Simpson 多样性指数显示出分离株之间的显著差异。

结论

参考株和野外分离株的 RAPD 图谱似乎没有随时间变化,这可能反映了样本中基因没有发生 DNA 突变。最近的野外分离株与参考株具有不同的 WCP 裂解物图谱,可能是因为定型株的传代次数可能会影响其蛋白质表达。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/34f7/3499290/08552d04397f/1471-2180-12-108-1.jpg

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