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Journal of Clinical Microbiology, August 2000, p. 3072-3075, Vol. 38, No. 8
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Genetic Characterization of Toxin A-Negative, Toxin B-Positive Clostridium difficile Isolates by PCR

J. Scott Moncrief,* Limin Zheng, Laurie M. Neville, and David M. Lyerly

TechLab, Inc., Blacksburg, Virginia 24060-6364

Received 8 February 2000/Returned for modification 5 May 2000/Accepted 17 May 2000

Toxin-specific enzyme immunoassays, cytotoxicity assays, and PCR were used to analyze 48 toxin A-negative, toxin B-positive Clostridium difficile isolates from various geographical sites around the world. All the isolates were negative by the TOX-A TEST and positive by the TOX A/B TEST. A deletion of approximately 1.7 kb was found at the 3' end of the toxA gene for all the isolates, similar to the deletion in toxinotype VIII strains (e.g., C. difficile serotype F 1470). Additional PCR analysis indicated that the toxin B encoded by these isolates contains sequence variations downstream of the active site compared to the sequence of reference strain VPI 10463. This variation may extend the glucosylation spectrum to Ras proteins, as observed previously for closely related lethal toxin from Clostridium sordellii and toxin B from toxin A-negative, toxin B-positive strain F 1470. Toxin A-negative, toxin B-positive isolates have recently been associated with disease in humans, and they may be more common than was previously supposed.

* Corresponding author. Mailing address: TechLab, Inc., VPI Research Park, 1861 Pratt Dr., Suite 1030, Blacksburg, VA 24060-6364. Phone: (540) 953-1664. Fax: (540) 953-1665. E-mail: jsmoncrief{at}techlabinc.com.

Journal of Clinical Microbiology, August 2000, p. 3072-3075, Vol. 38, No. 8
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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