HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Kato, H. Articles by Arakawa, Y. Search for Related Content PubMed PubMed Citation Articles by Kato, H. Articles by Arakawa, Y. Agricola Articles by Kato, H. Articles by Arakawa, Y.

Typing by sequencing the slpA gene of Clostridium difficile strains causing multiple outbreaks in Japan

¹Department of Bacterial Pathogenesis and Infection Control, National Institute of Infectious Diseases, 4-7-1 Gakuen, Musashimurayama, Tokyo 208-0011, Japan ²Kumiai Hospital, 5-68 Ozinmachi, Takayama Gifu 506-8502, Japan

Correspondence Haru Kato cato{at}nih.go.jp

Received July 1, 2004
Accepted October 18, 2004

Previous reports have documented that a surface layer protein (SlpA) varies among Clostridium difficile isolates. The typing system by sequencing the variable region of the slpA gene was applied to typing C. difficile strains belonging to one PCR ribotype, type smz, which has been identified as frequently causing outbreaks in Japan. The PCR ribotype smz strains recovered from patients at different hospitals in Japan were examined. Among 10 type smz strains tested, three subtypes, smz-1, -2 and -3, were identified that differed from each other by one nucleotide. slpA sequence typing was also applied to direct typing on DNA extracted from stool specimens. Of 22 stool specimens examined, 17 were PCR positive for slpA; eight were typed as slpA sequence type smz-1 and nine as type smz-2. C. difficile was cultured from 12 of these 17 stool specimens, and the sequence results of the recovered isolates were compared with those from the DNA extracted from the stool specimens. In all 12 of these stool specimens, the sequence results of DNA from recovered C. difficile isolates completely agreed with those of DNA extracted directly from stool specimens. The remaining five stool specimens were culture-negative for C. difficile. Sequence typing has the advantage of enabling easy comparison of typing results among multiple laboratories via the Internet without exchanging reference strains as is required in typing systems which depend on banding-pattern analyses. slpA sequence typing appears to be a reproducible and reliable typing system for C. difficile as well as being useful for the typing of C. difficile when stool specimens contain only small numbers of C. difficile or are inappropriate for culturing.

Abbreviations: AP-PCR, arbitrarily primed PCR; MLST, multilocus sequence typing; REA, restriction enzyme analysis.

The GenBank/EMBL/DDBJ accession numbers for the sequences of the variable regions of the slpA gene of strain GAI 97660 (slpA sequence type smz-1), strain MRY 04-0409 (slpA sequence type smz-2) and strain MRY 04-0410 (slpA sequence type smz-3) are AB180242, AB181350 and AB181351, respectively.

This article has been cited by other articles:

				G. Killgore, A. Thompson, S. Johnson, J. Brazier, E. Kuijper, J. Pepin, E. H. Frost, P. Savelkoul, B. Nicholson, R. J. van den Berg, et al. Comparison of Seven Techniques for Typing International Epidemic Strains of Clostridium difficile: Restriction Endonuclease Analysis, Pulsed-Field Gel Electrophoresis, PCR-Ribotyping, Multilocus Sequence Typing, Multilocus Variable-Number Tandem-Repeat Analysis, Amplified Fragment Length Polymorphism, and Surface Layer Protein A Gene Sequence Typing J. Clin. Microbiol., February 1, 2008; 46(2): 431 - 437. [Abstract] [Full Text] [PDF]

				I. Poilane, C. Humeniuk-Ainouz, I. Durand, C. Janoir, P. Cruaud, M. Delmee, M. R. Popoff, and A. Collignon Molecular characterization of Clostridium difficile clinical isolates in a geriatric hospital J. Med. Microbiol., March 1, 2007; 56(3): 386 - 390. [Abstract] [Full Text] [PDF]

				H. Kato, T. Yokoyama, H. Kato, and Y. Arakawa Rapid and Simple Method for Detecting the Toxin B Gene of Clostridium difficile in Stool Specimens by Loop-Mediated Isothermal Amplification J. Clin. Microbiol., December 1, 2005; 43(12): 6108 - 6112. [Abstract] [Full Text] [PDF]

				I. R Poxton Clostridium difficile J. Med. Microbiol., February 1, 2005; 54(2): 97 - 100. [Full Text] [PDF]