Rapid species identification and partial strain differentiation of Clostridium butyricum by PCR using 16S-23S rDNA intergenic spacer regions
Rapid species identification and partial strain differentiation of Clostridium butyricum by PCR using 16S-23S rDNA intergenic spacer regions
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DOI：
10.1111/j.1348-0421.2005.tb03653.x
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发表时间：
2005-01-01
影响因子：
2.6
通讯作者：
Ohnishi, Y
中科院分区：
文献类型：
--
作者：
Nakanishi, S;Kuwahara, T;Ohnishi, Y
一些丁酸梭菌菌株已被用作人和动物的益生菌。菌种特异性鉴定是益生菌生产过程中必不可少的环节。本研究的目的是确定是否有足够的遗传变异，在16 S-23 S基因间隔区（ISR）区分丁酸梭菌在生物群水平。我们从5个参考菌株，益生菌株（MIYAIRI 588）和22株分离株扩增的ISR，我们将它们分为四组的基础上的扩增模式（A型到D型）。然而，扩增的ISR是不足以区分菌株。此外，我们还比较了这些ISRs的遗传结构。序列分析表明，ISRs的大小变化是由tRNA基因和独特的序列插入到内部部分产生的，而外部部分是高度保守的。根据这两个ISRs高度保守的核苷酸序列，我们设计了一对特异性PCR引物。丁酸。此外，根据B型菌株中的独特插入序列设计的PCR引物可用于在生物群水平上区分益生菌菌株。
Some Clostridium butyricum strains have been used as probiotics for both humans and animals. Strain-specific identification is necessary for the manufacturing process of probiotics. The aim of this study was to determine whether there are sufficient genetic variations in 16S-23S intergenic spacer regions (ISRs) to discriminate C butyricum at the biovar level. We amplified ISRs from five reference strains, a probiotic strain (MIYAIRI 588) and 22 isolates, and we classified them into four groups on the basis of amplification patterns (type A through D). However, amplification of ISRs is not sufficient for discriminating strains. Moreover, we compared genetic structures of these ISRs. Sequence analysis revealed that the size variations of ISRs were generated by the insertion of tRNA genes and unique sequences into the internal portion, while the external portions were highly conserved. On the basis of the highly conserved nucleotide sequences within the ISRs, we developed a PCR primer set specific to C. butyricum. In addition, the PCR primer designed from the unique inserted sequence in type B strain was useful to differentiate probiotic strains at the biovar level.