利用聚合酶链反应扩增的16S rDNA的限制性片段长度多态性分析鉴别非典型韦荣球菌、殊异韦荣球菌和小韦荣球菌。

Differentiation of Veillonella atypica, Veillonella dispar and Veillonella parvula using restricted fragment-length polymorphism analysis of 16S rDNA amplified by polymerase chain reaction.

作者信息

Sato T, Matsuyama J, Sato M, Hoshino E

机构信息

Department of Oral Microbiology, Niigata University School of Dentistry, Japan.

出版信息

Oral Microbiol Immunol. 1997 Dec;12(6):350-3. doi: 10.1111/j.1399-302x.1997.tb00737.x.

DOI:10.1111/j.1399-302x.1997.tb00737.x

PMID:9573809

Abstract

Veillonella atypica, Veillonella dispar and Veillonella parvula cannot be reliably distinguished by conventional phenotypic tests, including the API ZYM test. In this study, restricted fragment-length polymorphism (RFLP) analysis of 16S rDNA amplified by polymerase chain reaction (PCR) was used to generate restriction profiles of the type strains of V. atypica, V. dispar and V. parvula and 20 Veillonella strains isolated from oral sites. 16S rRNA gene sequences from isolated genomic DNA samples were amplified by PCR. PCR products were purified and characterized by single digestion with 13 restriction endonucleases. Among them, Mn/I was found to discriminate the respective reference strains, and the clinical isolates were assigned to one of the three species on the basis of their restriction profiles by digestion with Mn/I. Thus, RFLP analysis of PCR-amplified 16S rDNA, using Mn/I, is a rapid and reliable method for the differentiation of V. atypica, V. dispar and V. parvula.

摘要

非典型韦荣球菌、殊异韦荣球菌和微小韦荣球菌无法通过包括API ZYM试验在内的传统表型试验可靠地区分。在本研究中，利用聚合酶链反应（PCR）扩增16S rDNA的限制性片段长度多态性（RFLP）分析，对非典型韦荣球菌、殊异韦荣球菌和微小韦荣球菌的模式菌株以及从口腔部位分离出的20株韦荣球菌菌株进行限制性图谱分析。通过PCR扩增分离的基因组DNA样本中的16S rRNA基因序列。PCR产物经纯化后用13种限制性内切酶进行单酶切鉴定。其中，发现Mn/I可区分各自的参考菌株，临床分离株根据其经Mn/I酶切后的限制性图谱被归为这三个种之一。因此，使用Mn/I对PCR扩增的16S rDNA进行RFLP分析，是区分非典型韦荣球菌、殊异韦荣球菌和微小韦荣球菌的快速可靠方法。