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Ribosomal DNA Sequencing for Identification of Aerobic Gram-Positive Rods in the Clinical Laboratory (an 18-Month Evaluation)

机译:核糖体DNA测序在临床实验室中鉴定需氧革兰阳性棒(18个月评估)

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摘要

We have evaluated over a period of 18 months the use of 16S ribosomal DNA (rDNA) sequence analysis as a means of identifying aerobic gram-positive rods in the clinical laboratory. Two collections of strains were studied: (i) 37 clinical strains of gram-positive rods well identified by phenotypic tests, and (ii) 136 clinical isolates difficult to identify by standard microbiological investigations, i.e., identification at the species level was impossible. Results of molecular analyses were compared with those of conventional phenotypic identification procedures. Good overall agreement between phenotypic and molecular identification procedures was found for the collection of 37 clinical strains well identified by conventional means. For the 136 clinical strains which were difficult to identify by standard microbiological investigations, phenotypic characterization identified 71 of 136 (52.2%) isolates at the genus level; 65 of 136 (47.8%) isolates could not be discriminated at any taxonomic level. In comparison, 16S rDNA sequencing identified 89 of 136 (65.4%) isolates at the species level, 43 of 136 (31.6%) isolates at the genus level, and 4 of 136 (2.9%) isolates at the family level. We conclude that (i) rDNA sequencing is an effective means for the identification of aerobic gram-positive rods which are difficult to identify by conventional techniques, and (ii) molecular identification procedures are not required for isolates well identified by phenotypic investigations.
机译:我们已经在18个月的时间内评估了16S核糖体DNA(rDNA)序列分析在临床实验室中鉴定有氧革兰氏阳性棒的方法的使用。研究了两套菌株:(i)37种临床表现良好的革兰氏阳性菌菌株,通过表型测试可以很好地鉴定出来;(ii)136种临床分离株很难通过标准微生物学调查来鉴定,即无法在物种水平上鉴定。分子分析的结果与常规表型鉴定程序进行了比较。表型和分子鉴定程序之间的良好总体协议被发现,以收集通过常规方法很好鉴定的37种临床菌株。对于通过标准微生物学调查难以鉴定的136株临床菌株,表型特征鉴定了属水平的136株菌株中的71株(占52.2%)。在任何分类学水平上,均不能区分136个菌株中的65个(47.8%)。相比之下,16S rDNA测序在物种水平上鉴定出136个菌株中的89个(65.4%),在属水平上鉴定了136个菌株中的43个(31.6%),在家族水平上鉴定了136个菌株中的4个(2.9%)。我们得出的结论是:(i)rDNA测序是鉴定需氧革兰氏阳性棒的有效方法,而传统技术难以鉴定,并且(ii)对于通过表型研究很好鉴定的分离株,不需要分子鉴定程序。

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