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首页> 外文期刊>Journal of Clinical Microbiology >Polymerase chain reaction-based restriction fragment length polymorphism analysis of a fragment of the ribosomal operon from Rochalimaea species for subtyping.
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Polymerase chain reaction-based restriction fragment length polymorphism analysis of a fragment of the ribosomal operon from Rochalimaea species for subtyping.

机译：基于聚合酶链反应的限制性内切酶片段长度多态性分析，用于亚克隆的罗莎木属物种核糖体操纵子片段。

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Restriction endonuclease analysis of a polymerase chain reaction-amplified DNA fragment which included the spacer region between the genes coding for 16S and 23S rRNAs and a portion of the gene coding for 23S rRNA (spacer + 23S) was done on 10 previously characterized clinical isolates of Rochalimaea henselae, one clinical isolate of Rochalimaea quintana, and the type strains of R. henselae, R. quintana, Rochalimaea vinsonii, and Bartonella bacilliformis. Brucella abortus DNA was not amplified by the primer set used. The clinical isolates of Rochalimaea were obtained from blood or tissue from patients with and without preexisting disease. The amplicon from each strain was digested with five endonucleases (AluI, HaeIII, TaqI, HinfI, and MseI). AluI and HaeIII were useful in species differentiation and subtyping of R. henselae. R. henselae isolates showed six different restriction patterns with AluI and four patterns with HaeIII. TaqI, HinfI, and MseI were useful only in species differentiation. These observations indicate that PCR amplification of the spacer + 23S region of the ribosomal DNA of Rochalimaea spp., along with restriction endonuclease analysis, allows differentiation of Rochalimaea spp. from closely related genera, differentiation among the species within Rochalimaea, and differentiation of strains within R. henselae. The subtyping potential of this method may be useful for further clinical and epidemiologic studies of the spectrum of diseases caused by R. henselae.
机译：对聚合酶链反应扩增的DNA片段进行了限制性核酸内切酶分析，该片段包括对16S和23S rRNA编码的基因之间的间隔区和对23S rRNA编码的一部分基因（间隔区+ 23S），对10个先前鉴定的Hs分离株进行了分析。罗汉木（Rochalimaea henselae），罗汉木（Rochalimaea quintana）的一种临床分离株，以及罗汉木，罗汉木（R. quintana），罗汉木（Rochalimaea vinsonii）和杆菌Bartonella bacilliformis的类型株。使用的引物组不能扩增流产布鲁氏菌DNA。 Rochalimaea的临床分离株是从患有或未患有已有疾病的患者的血液或组织中获得的。用五种核酸内切酶（AluI，HaeIII，TaqI，HinfI和MseI）消化每个菌株的扩增子。 AluI和HaeIII可用于亨氏梭菌的物种分化和亚型化。 R. henselae分离株对AluI表现出六种不同的限制模式，对HaeIII表现出四种模式。 TaqI，HinfI和MseI仅在物种分化中有用。这些观察结果表明，Rochalimaea spp。的核糖体DNA的间隔区+ 23S区域的PCR扩增以及限制性核酸内切酶分析允许Rochalimaea spp的分化。来自密切相关的属，在罗氏藻内的种间分化，以及在汉斯酵母中的菌株分化。这种方法的亚型化潜能可能对进一步研究由汉森氏杆菌引起的疾病的临床和流行病学研究有用。

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