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通过扩增片段长度多态性分析、脉冲场凝胶电泳和16S核糖体DNA测序对猪滑膜支原体菌株进行鉴定

Characterization of Mycoplasma hyosynoviae strains by amplified fragment length polymorphism analysis, pulsed-field gel electrophoresis and 16S ribosomal DNA sequencing.

作者信息

Kokotovic B, Friis N F, Ahrens P

机构信息

Danish Veterinary Laboratory, Copenhagen.

出版信息

J Vet Med B Infect Dis Vet Public Health. 2002 Jun;49(5):245-52. doi: 10.1046/j.1439-0450.2002.00558.x.

Abstract

Mycoplasma hyospnoviae strains from Denmark, Germany, Japan, Sweden, the Netherlands and the UK were examined for variations in the genomic DNA and within the 16S ribosomal RNA (rRNA) gene. Variations in the chromosomal DNA among 57 isolates recovered from the respiratory tract and joints of pigs, were investigated by analysis of amplified fragment length polymorphisms of the Bg/II and MfeI restriction sites and by pulsed-field gel electrophoresis of a BssHII digest of chromosomal DNA. Both methods allowed unambiguous differentiation of the analysed strains and showed similar discriminatory potential for the differentiation of M. hyosynoviae isolates. Concordant results obtained with the two whole-genome fingerprinting techniques evidence the considerable intraspecies genetic heterogeneity of M. hyosynoviae. Sixteen field strains of M. hyosynoviae and the type strain S16(T) were further examined for variation within the 16S rRNA gene. Ten field strains possessed the 16S rDNA sequences identical to the type strain, while the remaining six strains had sequences that differed by one to two nucleotides from that obtained from the type strain.

摘要

对来自丹麦、德国、日本、瑞典、荷兰和英国的猪肺炎支原体菌株进行了基因组DNA和16S核糖体RNA(rRNA)基因变异的检测。通过分析Bg/II和MfeI限制性位点的扩增片段长度多态性以及对染色体DNA的BssHII酶切产物进行脉冲场凝胶电泳,研究了从猪的呼吸道和关节中分离出的57株菌株的染色体DNA变异情况。两种方法都能明确区分所分析的菌株,并且对猪肺炎支原体分离株的鉴别具有相似的区分潜力。两种全基因组指纹技术获得的一致结果证明了猪肺炎支原体在种内存在相当大的遗传异质性。进一步检测了16株猪肺炎支原体田间菌株和模式菌株S16(T)的16S rRNA基因变异情况。10株田间菌株的16S rDNA序列与模式菌株相同,而其余6株菌株的序列与模式菌株相比有1至2个核苷酸的差异。

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