DIFFERENTIATION OF STREPTOCOCCUS-UBERIS FROM STREPTOCOCCUS-PARAUBERIS BY POLYMERASE CHAIN-REACTION AND RESTRICTION-FRAGMENT-LENGTH-POLYMORPHISM ANALYSIS OF 16S RIBOSOMAL DNA
DIFFERENTIATION OF STREPTOCOCCUS-UBERIS FROM STREPTOCOCCUS-PARAUBERIS BY POLYMERASE CHAIN-REACTION AND RESTRICTION-FRAGMENT-LENGTH-POLYMORPHISM ANALYSIS OF 16S RIBOSOMAL DNA
复制标题
DOI：
10.1128/jcm.29.12.2774-2778.1991
复制
发表时间：
1991-12-01
影响因子：
9.4
通讯作者：
OLIVER, SP
中科院分区：
文献类型：
--
作者：
JAYARAO, BM;DORE, JJE;OLIVER, SP
Uberis type II是最近提出的一个单独的种，命名为副链球菌(A.M.Williams和M.D.Collins，J.Appl.)细菌素。68：485-490,1990)。由于副葡萄球菌和Uberis的生化和血清学特征无法区分，因此只能通过DNA-DNA杂交或16S rRNA测序才能区分这两个物种。通过对16S核糖体DNA 1.4kb的限制性片段长度多态性分析，建立了一种简单可靠的鉴别副葡萄球菌II型[ATCC 13386]和Uberis I型[ATCC9927、ATCC 13387和ATCC 27958]的方法。利用与16S rRNA基因互补的寡核苷酸引物，通过聚合酶链式反应从基因组DNA中扩增出16S核糖体基因片段。用SCAI、NSPI、DdeI和AvaII限制性内切酶对1.4kb的16S rDNA片段进行酶切。所有四种限制性内切酶产生的限制性内切酶片段都是每个物种特有的。对从奶牛乳腺分泌物中分离到的24个“Uberis”菌株的16S rDNA进行RFLP分析，结果表明，所有24个菌株均为S.uberis。
Streptococcus uberis type II has been proposed recently as a separate species designated Streptococcus parauberis (A. M. Williams and M. D. Collins, J. Appl. Bacteriol. 68:485-490, 1990). Differentiation of S. parauberis from S. uberis has been possible only by DNA-DNA hybridization or 16S rRNA sequencing, since the biochemical and serological characteristics of the two species are indistinguishable. A simple and reliable technique was developed for differentiating S. parauberis (S. uberis type II [ATCC 13386]) from S. uberis (S. uberis type I [ATCC 9927, ATCC 13387, and ATCC 27958]) by restriction fragment length polymorphism (RFLP) analysis of 1.4-kb 16S ribosomal DNA (rDNA). Oligonucleotide primers complementary to 16S rRNA genes were used to amplify 16S ribosomal gene fragments from genomic DNA by polymerase chain reaction. The 1.4-kb 16S rDNA fragment was digested with ScaI, NspI, DdeI, and AvaII restriction endonucleases. Restriction fragments produced by all four restriction endonucleases were characteristic for each species. RFLP analysis of 16S rDNA from 24 "S. uberis" isolates obtained from mammary secretions of dairy cows indicated that all 24 isolates were indeed S. uberis.