Biotechnical Faculty, Institute of Dairy Science and Probiotics, University of Ljubljana, Groblje 3, 1230, Domžale, Slovenia,
Folia Microbiol (Praha). 2013 Nov;58(6):623-30. doi: 10.1007/s12223-013-0252-8. Epub 2013 Apr 25.
Lactobacillus gasseri K7 is a probiotic strain that produces bacteriocins gassericin K7 A and K7 B. In order to develop a real-time quantitative PCR assay for the detection of L. gasseri K7, 18 reference strains of the Lactobacillus acidophilus group and 45 faecal samples of adults who have never consumed strain K7 were tested with PCR using 14 pairs of primers specific for gassericin K7 A and K7 B gene determinants. Incomplete gassericin K7 A or K7 B gene clusters were found to be dispersed in different lactobacilli strains as well as in faecal microbiota. One pair of primers was found to be specific for the total gene cluster of gassericin K7A and one for gassericin K7B. The real-time PCR analysis of faecal samples spiked with K7 strain revealed that primers specific for the gene cluster of the gassericin K7 A were more suitable for quantitative determination than those for gassericin K7 B, due to the lower detection level. Targeting of the gassericin K7 A or K7 B gene cluster with specific primers could be used for detection and quantification of L. gasseri K7 in human faecal samples without prior cultivation. The results of this study also present new insights into the prevalence of bacteriocin-encoding genes in gastrointestinal tract.
格氏乳杆菌 K7 是一种益生菌菌株,可产生细菌素 gassericin K7A 和 K7B。为了开发用于检测 L. gasseri K7 的实时定量 PCR 检测方法,使用针对 gassericin K7A 和 K7B 基因决定簇的 14 对引物对 18 株嗜酸乳杆菌属参考菌株和 45 份从未食用过 K7 菌株的成人粪便样本进行了 PCR 测试。发现不完全的 gassericin K7A 或 K7B 基因簇分散在不同的乳杆菌菌株以及粪便微生物群中。发现一对引物特异性针对 gassericin K7A 的总基因簇,另一对引物特异性针对 gassericin K7B。用 K7 菌株接种的粪便样本的实时 PCR 分析表明,由于检测水平较低,针对 gassericin K7A 基因簇的引物比针对 gassericin K7B 的引物更适合定量测定。使用特异性引物针对 gassericin K7A 或 K7B 基因簇进行靶向检测和定量可用于检测和定量人粪便样本中的 L. gasseri K7,而无需事先培养。本研究的结果还提供了关于胃肠道中细菌素编码基因流行率的新见解。