Evaluation of tRNA Gene PCR for Identification of Mollicutes

Tim Stakenborg; Jo Vicca; Rita Verhelst; Patrick Butaye; Dominiek Maes; Anne Naessens; Geert Claeys; Catharine De Ganck; Freddy Haesebrouck; Mario Vaneechoutte

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Evaluation of tRNA Gene PCR for Identification of Mollicutes

机译：评价tRNA基因PCR鉴定囊肿的方法。

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We evaluated the applicability of tRNA gene PCR in combination with fluorescent capillary electrophoresis with an ABI310 genetic analyzer (Applied Biosystems, Calif.) for the identification of different mollicute species. A total of 103 strains and DNA extracts of 30 different species belonging to the genera Acholeplasma, Mycoplasma, and Ureaplasma were studied. Reproducible peak profiles were generated for all samples, except for one M. genitalium isolate, the three M. gallisepticum isolates, and 8 of the 24 Ureaplasma cultures, where no amplification could be obtained. Clustering revealed numerous discrepancies compared to the identifications that had been previously obtained by means of biochemical and serological tests. Final identification was obtained by 16S rRNA gene amplification followed by sequence analysis and/or restriction digestion. This confirmed the identification obtained by tRNA gene PCR in all cases. Seven samples yielded an unexpected tRNA gene PCR profile. Sequence analysis of the 16S rRNA genes showed that six of these samples were mixed and that one had a unique sequence that did not match any of the published sequences, pointing to the existence of a not-yet-described species. In conclusion, we found tRNA gene PCR to be a rapid and discriminatory method to correctly identify a large collection of different species of the class of Mollicutes and to recognize not-yet-described groups.

机译：我们评估了tRNA基因PCR与荧光毛细管电泳结合ABI310基因分析仪（Applied Biosystems，CA）的适用性，以鉴定不同的分子种类。研究了共计103种菌株，分别属于 Acholeplasma ， Mycoplasma 和 Ureaplasma 的30个不同物种。除一个 M以外，所有样品均产生了可再现的峰图。生殖器分离物，三个 M。鸡败血杆菌分离株，以及24种 Ureaplasma 培养物中的8种，无法扩增。与以前通过生化和血清学测试获得的鉴定结果相比，聚类显示出许多差异。通过16S rRNA基因扩增，序列分析和/或限制性酶切消化获得最终鉴定。这证实了在所有情况下通过tRNA基因PCR获得的鉴定。七个样品产生了意外的tRNA基因PCR谱。对16S rRNA基因的序列分析表明，这些样品中有6个是混合的，其中一个的独特序列与任何已公开的序列都不匹配，这说明该物种尚未存在。总之，我们发现tRNA基因PCR是一种快速而有区别的方法，可以正确地识别大量的 Mollicutes 物种并识别尚未描述的群体。 展开▼

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《Journal of Clinical Microbiology》 |2005年第9期|共9页

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Tim Stakenborg; Jo Vicca; Rita Verhelst; Patrick Butaye; Dominiek Maes; Anne Naessens; Geert Claeys; Catharine De Ganck; Freddy Haesebrouck; Mario Vaneechoutte;
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中图分类医学微生物学（病原细菌学、病原微生物学）;

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2. Species-Specific PCR for Identification of Common Contaminant Mollicutes in Cell Culture [J] . Fanrong Kong, Gregory James, Susanna Gordon, Applied and Environmental Microbiology . 2001,第7期

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6. Evaluation of tRNA Gene PCR for Identification of Mollicutes [O] . Tim Stakenborg, Jo Vicca, Rita Verhelst, 2018

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7. Evaluation of tRNA gene PCR for identification of Mollicutes [O] . Stakenborg T, Vicca Jo, Verhelst R, 2005

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Evaluation of tRNA Gene PCR for Identification of Mollicutes

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