Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
Comparative Study
. 1999 Sep;65(9):3908-14.
doi: 10.1128/AEM.65.9.3908-3914.1999.

Comparison of ribotyping, randomly amplified polymorphic DNA analysis, and pulsed-field gel electrophoresis in typing of Lactobacillus rhamnosus and L. casei strains

S Tynkkynen  1 , R Satokari, M Saarela, T Mattila-Sandholm, M Saxelin
Affiliations
Free PMC article
Comparative Study

Comparison of ribotyping, randomly amplified polymorphic DNA analysis, and pulsed-field gel electrophoresis in typing of Lactobacillus rhamnosus and L. casei strains

S Tynkkynen et al. Appl Environ Microbiol. 1999 Sep.
Free PMC article

Abstract

A total of 24 strains, biochemically identified as members of the Lactobacillus casei group, were identified by PCR with species-specific primers. The same set of strains was typed by randomly amplified polymorphic DNA (RAPD) analysis, ribotyping, and pulsed-field gel electrophoresis (PFGE) in order to compare the discriminatory power of the methods. Species-specific primers for L. rhamnosus and L. casei identified the type strain L. rhamnosus ATCC 7469 and the neotype strain L. casei ATCC 334, respectively, but did not give any signal with the recently revived species L. zeae, which contains the type strain ATCC 15820 and the strain ATCC 393, which was previously classified as L. casei. Our results are in accordance with the suggested new classification of the L. casei group. Altogether, 21 of the 24 strains studied were identified with the species-specific primers. In strain typing, PFGE was the most discriminatory method, revealing 17 genotypes for the 24 strains studied. Ribotyping and RAPD analysis yielded 15 and 12 genotypes, respectively.

PubMed Disclaimer

Figures

FIG. 1
FIG. 1
RAPD patterns and genotypes (in parentheses) of the strains. Lanes: 1 to 18, L. rhamnosus GG (A1), VS 1031 (A1), VS 1032 (A1), VS 1034 (A1), VS 1017 (A1), VS 1018 (A1), ATCC 7469 (A2), ATCC 11443 (A2), E-78080 (A2), VS 872 (A2), VS 495 (A2), VS 1022 (A2), VS 1020 (A3), VS 1021 (A3), E-97800 (A4), VS 1030 (A5), Lactophilus (A6), and VS 1019 (A5), respectively; 19, L. casei VS 1023 (A7); 20, L. rhamnosus VS 1033 (A8); 21, L. zeae ATCC 15820 (A9); 22, L. casei ATCC 393 (A10); 23, L. casei ATCC 334 (A11); 24, L. casei ATCC 4646 (A12); 25, molecular weight marker (in kilobase pairs).
FIG. 2
FIG. 2
RiboPrint patterns of L. rhamnosus, L. casei, and L. zeae strains. The patterns are composites of several individual patterns. Ribotypes: R1, L. rhamnosus GG, VS 1032, VS 1034, VS 1018, VS 1031, and VS 1017; R2, L. rhamnosus ATCC 7469, ATCC 11443, and E-78080; R3, L. rhamnosus VS 872 and VS 1022; R4, L. rhamnosus VS 495; R5, L. rhamnosus VS 1020; R6, L. rhamnosus VS 1021; R7, L. rhamnosus E-97800 and VS 1019; R8, L. rhamnosus VS 1030; R9, L. rhamnosus Lactophilus; R10, L. rhamnosus VS 1033; R11, L. casei VS 1023; R12, L. casei ATCC 393; R13, L. casei ATCC 334; R14, L. casei ATCC 4646; R15, L. zeae ATCC 15820.
FIG. 3
FIG. 3
PFGE profiles and genotypes (in parentheses) of the strains as determined with restriction enzyme SfiI. (a) Lanes: 1 to 9, L. rhamnosus GG (S1), ATCC 7469 (S2), ATCC 11443 (S2), E-78080 (S2), VS 872 (S2), E-97800 (S3), VS 1030 (S4), VS 1033 (S5), and VS 1021 (S6), respectively; 10, L. zeae ATCC 15820 (S7). (b) Lanes: 1 to 10, L. rhamnosus VS 1031 (S8), VS 1032 (S1), VS 1034 (S1), Lactophilus (S9), VS 495 (S10), VS 1017 (S11), VS 1018 (S1), VS 1019 (S12), VS 1020 (S6), and VS 1022 (S2), respectively. (c) Lanes: 1 to 4, L. casei VS 1023 (S13), ATCC 393 (S14), ATCC 334 (S15), and ATCC 4646 (S16), respectively.
FIG. 3
FIG. 3
PFGE profiles and genotypes (in parentheses) of the strains as determined with restriction enzyme SfiI. (a) Lanes: 1 to 9, L. rhamnosus GG (S1), ATCC 7469 (S2), ATCC 11443 (S2), E-78080 (S2), VS 872 (S2), E-97800 (S3), VS 1030 (S4), VS 1033 (S5), and VS 1021 (S6), respectively; 10, L. zeae ATCC 15820 (S7). (b) Lanes: 1 to 10, L. rhamnosus VS 1031 (S8), VS 1032 (S1), VS 1034 (S1), Lactophilus (S9), VS 495 (S10), VS 1017 (S11), VS 1018 (S1), VS 1019 (S12), VS 1020 (S6), and VS 1022 (S2), respectively. (c) Lanes: 1 to 4, L. casei VS 1023 (S13), ATCC 393 (S14), ATCC 334 (S15), and ATCC 4646 (S16), respectively.
FIG. 3
FIG. 3
PFGE profiles and genotypes (in parentheses) of the strains as determined with restriction enzyme SfiI. (a) Lanes: 1 to 9, L. rhamnosus GG (S1), ATCC 7469 (S2), ATCC 11443 (S2), E-78080 (S2), VS 872 (S2), E-97800 (S3), VS 1030 (S4), VS 1033 (S5), and VS 1021 (S6), respectively; 10, L. zeae ATCC 15820 (S7). (b) Lanes: 1 to 10, L. rhamnosus VS 1031 (S8), VS 1032 (S1), VS 1034 (S1), Lactophilus (S9), VS 495 (S10), VS 1017 (S11), VS 1018 (S1), VS 1019 (S12), VS 1020 (S6), and VS 1022 (S2), respectively. (c) Lanes: 1 to 4, L. casei VS 1023 (S13), ATCC 393 (S14), ATCC 334 (S15), and ATCC 4646 (S16), respectively.
FIG. 4
FIG. 4
PFGE profiles and genotypes (in parentheses) of the strains as determined with restriction enzyme NotI. (a) Lanes: 1 to 9, L. rhamnosus GG (N1), ATCC 7469 (N2), ATCC 11443 (N2), E-78080 (N2), VS 872 (N3), E-97800 (N4), VS 1030 (N5), VS 1033 (N6), and VS 1021 (N7), respectively; 10, L. zeae ATCC 15820 (N8). (b) Lanes: 1 to 10, L. rhamnosus VS 1031 (N1), VS 1032 (N1), VS 1034 (N1), Lactophilus (N9), VS 495 (N10), VS 1017 (N11), VS 1018 (N1), VS 1019 (N5), VS 1020 (N7), and VS 1022 (N3), respectively. (c) Lanes: 1 to 4, L. casei VS 1023 (N12), ATCC 393 (N13), ATCC 334 (N14), and ATCC 4646 (N15), respectively.
FIG. 4
FIG. 4
PFGE profiles and genotypes (in parentheses) of the strains as determined with restriction enzyme NotI. (a) Lanes: 1 to 9, L. rhamnosus GG (N1), ATCC 7469 (N2), ATCC 11443 (N2), E-78080 (N2), VS 872 (N3), E-97800 (N4), VS 1030 (N5), VS 1033 (N6), and VS 1021 (N7), respectively; 10, L. zeae ATCC 15820 (N8). (b) Lanes: 1 to 10, L. rhamnosus VS 1031 (N1), VS 1032 (N1), VS 1034 (N1), Lactophilus (N9), VS 495 (N10), VS 1017 (N11), VS 1018 (N1), VS 1019 (N5), VS 1020 (N7), and VS 1022 (N3), respectively. (c) Lanes: 1 to 4, L. casei VS 1023 (N12), ATCC 393 (N13), ATCC 334 (N14), and ATCC 4646 (N15), respectively.
FIG. 4
FIG. 4
PFGE profiles and genotypes (in parentheses) of the strains as determined with restriction enzyme NotI. (a) Lanes: 1 to 9, L. rhamnosus GG (N1), ATCC 7469 (N2), ATCC 11443 (N2), E-78080 (N2), VS 872 (N3), E-97800 (N4), VS 1030 (N5), VS 1033 (N6), and VS 1021 (N7), respectively; 10, L. zeae ATCC 15820 (N8). (b) Lanes: 1 to 10, L. rhamnosus VS 1031 (N1), VS 1032 (N1), VS 1034 (N1), Lactophilus (N9), VS 495 (N10), VS 1017 (N11), VS 1018 (N1), VS 1019 (N5), VS 1020 (N7), and VS 1022 (N3), respectively. (c) Lanes: 1 to 4, L. casei VS 1023 (N12), ATCC 393 (N13), ATCC 334 (N14), and ATCC 4646 (N15), respectively.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Alander M, Satokari R, Korpela R, Saxelin M, Vilpponen-Salmela T, Mattila-Sandholm T, von Wright A. Persistence of colonization of human colonic mucosa by a probiotic strain, Lactobacillus rhamnosus GG, after oral consumption. Appl Environ Microbiol. 1999;65:351–354. - PMC - PubMed
    1. Barry T, Colleran G, Glennon M, Dunican L K, Gannon F. The 16S/23S ribosomal spacer region as a target for DNA probes to identify eubacteria. PCR Methods Appl. 1991;1:51–56. - PubMed
    1. Björkroth J, Ridell J, Korkeala H. Characterization of Lactobacillus sake strains associating with production of ropy slime by randomly amplified polymorphic DNA (RAPD) and pulsed-field gel electrophoresis (PFGE) patterns. Int J Food Microbiol. 1996;31:59–68. - PubMed
    1. Bollet C, Gevaudan M J, de Lamballerie X, Zandotti C, de Micco P. A simple method for the isolation of chromosomal DNA from gram positive or acid fast bacteria. Nucleic Acids Res. 1991;19:1955. - PMC - PubMed
    1. Brosius J, Ullrich A, Raker M A, Gray A, Dull T J, Gutell R R, Noller H F. Construction and fine mapping of recombinant plasmids containing the rrnB ribosomal RNA operon of E. coli. Plasmid. 1981;6:112–118. - PubMed

Publication types

MeSH terms

LinkOut - more resources