通过聚合酶链反应检测耐酒精火落菌
Detection of alcohol-tolerant hiochi bacteria by PCR.
作者信息
Nakagawa T, Shimada M, Mukai H, Asada K, Kato I, Fujino K, Sato T
机构信息
Biotechnology Research Laboratories, Takara Shuzo Co., Ltd., Shiga, Japan.
出版信息
Appl Environ Microbiol. 1994 Feb;60(2):637-40. doi: 10.1128/aem.60.2.637-640.1994.
We report a sensitive and rapid method for detection of hiochi bacteria by PCR. This method involves the electrophoresis of amplified DNA. Nucleotide sequences of the spacer region between 16S and 23S rRNA genes of 11 Lactobacillus strains were identified by analysis of PCR products. Five primers were designed by analysis of similarities among these sequences. A single cell of Lactobacillus casei subsp. casei could be detected when purified genomic DNA was used as the template. When various cell concentrations of L. casei subsp. casei were added to 50 ml of pasteurized sake and the cells were recovered, the detection limit was about one cell. No discrete band was observed in electrophoresis after PCR when human, Escherichia coli, mycoplasma, Acholeplasma, yeast, or mold DNA was used as the template.
我们报告了一种通过聚合酶链反应（PCR）检测火落菌的灵敏且快速的方法。该方法涉及扩增DNA的电泳。通过对PCR产物的分析，鉴定了11株乳酸杆菌菌株16S和23S rRNA基因间间隔区的核苷酸序列。通过分析这些序列之间的相似性设计了5种引物。当使用纯化的基因组DNA作为模板时，可检测到干酪乳杆菌干酪亚种的单个细胞。当将不同细胞浓度的干酪乳杆菌干酪亚种添加到50毫升巴氏杀菌清酒中并回收细胞时，检测限约为一个细胞。当使用人、大肠杆菌、支原体、无胆甾原体、酵母或霉菌DNA作为模板时，PCR后电泳未观察到离散条带。
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