Rapid identification of clinically significant species and taxa of aerobic actinomycetes, including Actinomadura, Gordona, Nocardia, Rhodococcus, Streptomyces, and Tsukamurella isolates, by DNA amplification and restriction endonuclease analysis
Rapid identification of clinically significant species and taxa of aerobic actinomycetes, including Actinomadura, Gordona, Nocardia, Rhodococcus, Streptomyces, and Tsukamurella isolates, by DNA amplification and restriction endonuclease analysis
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DOI：
10.1128/jcm.35.4.817-822.1997
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发表时间：
1997-04-01
影响因子：
9.4
通讯作者：
Wallace, RJ
中科院分区：
文献类型：
--
作者：
Steingrube, VA;Wilson, RW;Wallace, RJ
先前描述的PCR-限制性片段长度多态性（RFLP）鉴定模式诺卡氏菌，使用扩增的439-bp片段（扩增子）的65-kDa的热休克蛋白基因进行了评估，潜在的使用与所有临床上显着的好氧放线菌菌株。该研究包括28个参考菌株（美国典型培养物保藏中心）和198个临床分离株，属于20个分类组。在这198个分离物中，188个可以通过这种PCR-RFLP方法区分。所有需氧放线菌分离株的扩增子缺乏BstEII识别位点，从而将它们与含有一个或多个此类位点的分枝杆菌区分开来。在29种限制性内切酶中，MspI加HinfI产生的RFLP模式区分了20个分类群中的16个。一个单一的RFLP模式，观察到15 20个类群，其中包括65%的表型聚类菌株。在支气管戈登氏菌、星状诺卡氏菌复合物VI型、奥蒂斯卡夫诺卡氏菌、transvalensis诺卡氏菌和链霉菌属中观察到多种模式。链霉菌的RFLP图谱最不均一（19个菌株中有5个图谱），但显示出一个独特的大于320 bp的HinfI片段。从19个链霉菌分离株中的14个中获得了与白色链霉菌、灰色链霉菌或索马里链霉菌的模式菌株相匹配的RFLP图谱。28株N. 188株临床分离株中只有6株（3.2%）表现出与21种RFLP模式不匹配的模式。这些研究扩展了使用PCR-RFLP分析作为鉴定所有临床上重要的需氧放线菌物种和分类群的快速方法的可行性。
A previously described PCR-restriction fragment length polymorphism (RFLP) identification schema for Nocardia that used an amplified 439-bp segment (amplicon) of the 65-kDa heat shock protein gene was evaluated for potential use with isolates of all clinically significant aerobic actinomycetes. The study included 28 reference (American Type Culture Collection) strains and 198 clinical isolates belonging to 20 taxonomic groups. Of these 198 isolates, 188 could be differentiated by this PCR-RFLP method. Amplicons from all aerobic actinomycete isolates lacked BstEII recognition sites, thereby distinguishing them from those of mycobacteria that contain one or more such sites. Of 29 restriction endonucleases, MspI plus HinfI produced RFLP patterns that differentiated 16 of the 20 taxa. A single RFLP pattern was observed for 15 of 20 taxa that included 65% of phenotypically clustered isolates. Multiple patterns were seen with Gordona bronchialis, Nocardia asteroides complex type VI, Nocardia otitidiscaviarum, Nocardia transvalensis, and Streptomyces spp. Streptomyces RFLP patterns were the most heterogeneous (five patterns among 19 isolates), but exhibited a unique HinfI fragment of >320 bp. RFLP patterns that matched those from type strains of Streptomyces albus, Streptomyces griseus, or Streptomyces somaliensis were obtained from 14 of 19 Streptomyces isolates. Only 10 of 28 isolates of N. otitidiscaviarum failed to yield satisfactory amplicons, while only 6 of 188 (3.2%) clinical isolates exhibited patterns that failed to match one of the 21 defined RFLP patterns. These studies extended the feasibility of using PCR-RFLP analysis as a rapid method for the identification of all clinically significant species and taxa of aerobic actinomycetes.