Detection of <i>Lactobacillus, Pediococcus, Leuconostoc</i> , and <i>Weissella</i> Species in Human Feces by Using Group-Specific PCR Primers and Denaturing Gradient Gel Electrophoresis

Applied and Environmental Microbiology - Tập 67 Số 6 - Trang 2578-2585 - 2001
Jens Walter1, Christian Hertel1, Gerald W. Tannock2, Claudia M. Lis1, Karen Munro2, Walter P. Hammes1
1Institute of Food Technology, University of Hohenheim, Stuttgart, Germany,1 and
2Department of Microbiology, University of Otago, Dunedin, New Zealand2

Tóm tắt

ABSTRACT Denaturing gradient gel electrophoresis (DGGE) of DNA fragments generated by PCR with 16S ribosomal DNA-targeted group-specific primers was used to detect lactic acid bacteria (LAB) of the genera Lactobacillus, Pediococcus, Leuconostoc , and Weissella in human feces. Analysis of fecal samples of four subjects revealed individual profiles of DNA fragments originating not only from species that have been described as intestinal inhabitants but also from characteristically food-associated bacteria such as Lactobacillus sakei, Lactobacillus curvatus, Leuconostoc mesenteroides , and Pediococcus pentosaceus . Comparison of PCR-DGGE results with those of bacteriological culture showed that the food-associated species could not be cultured from the fecal samples by plating on Rogosa agar. On the other hand, all of the LAB species cultured from feces were detected in the DGGE profile. We also detected changes in the types of LAB present in human feces during consumption of a milk product containing the probiotic strain Lactobacillus rhamnosus DR20. The analysis of fecal samples from two subjects taken before, during, and after administration of the probiotic revealed that L. rhamnosus was detectable by PCR-DGGE during the test period in the feces of both subjects, whereas it was detectable by culture in only one of the subjects.

Từ khóa


Tài liệu tham khảo

10.1016/S0022-2836(05)80360-2

10.1128/AEM.66.9.3664-3673.2000

10.1016/0022-2836(81)90508-8

10.1136/gut.47.1.79

Hammes W. P. Vogel R. F. The genus Lactobacillus The lactic acid bacteria Wood B. J. B. Holzapfel W. H. 2. The genera of lactic acid bacteria 1995 19 54 Blackie Academic and Professional London, United Kingdom

10.1128/aem.63.8.3233-3241.1997

10.1128/JB.180.18.4765-4774.1998

10.1016/S0168-1605(98)00049-X

10.1128/AEM.65.10.4506-4512.1999

10.1128/aem.62.12.4608-4613.1996

10.1093/ajcn/71.4.861

Mitsuoka T. The human gastrointestinal tract The lactic acid bacteria Wood B. J. B. 1. The lactic acid bacteria in health and disease 1992 69 114 Elsevier Applied Science London, United Kingdom

10.1099/00207713-47-1-54

10.1023/A:1000669317571

10.1128/aem.63.8.3327-3332.1997

O'Sullivan D. Methods for analysis of the intestinal microflora Probiotics: a critical review. Tannock G. W. 1999 23 44 Horizon Scientific Press Wymondham United Kingdom

10.1128/AEM.65.9.3763-3766.1999

10.1016/S0723-2020(99)80039-3

10.1128/AEM.66.5.2263-2266.2000

10.1128/AEM.66.11.4705-4714.2000

10.1016/S0168-1605(96)01233-0

Tannock G. W. Normal microflora. An introduction to microbes inhabiting the human body. 1995 Chapman and Hall London United Kingdom

10.1023/A:1002038308506

10.1128/AEM.66.6.2578-2588.2000

10.1016/S0958-1669(99)00018-X

10.1128/AEM.66.1.297-303.2000

10.1128/aem.62.4.1242-1247.1996

Wu Y. Hayes V. M. Osinga J. Mulder I. M. Looman M. W. G. Buys C. H. C. M. Hofstra R. M. W. Improvement of fragment and primer selection for mutation detection by denaturing gradient gel electrophoresis. Nucleic Acids Res. 54 1998 32 40

10.1128/AEM.64.10.3854-3859.1998