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Infection and Immunity, December 1998, p. 5819-5825, Vol. 66, No. 12
Molecular Genetics Laboratory, International
Centre for Diarrhoeal Disease Research, Bangladesh, Dhaka-1000,
Bangladesh,1 and
Department of
Microbiology and Molecular Genetics, Harvard Medical School,
Boston, Massachusetts 021152
Received 29 June 1998/Returned for modification 19 August
1998/Accepted 5 September 1998
Toxigenic Vibrio cholerae strains are lysogens of
CTX
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Analysis of Clinical and Environmental Strains of
Nontoxigenic Vibrio cholerae for Susceptibility to CTX
:
Molecular Basis for Origination of New Strains with Epidemic
Potential
, a filamentous phage which encodes cholera toxin. The receptor
for CTX
for invading V. cholerae cells is the
toxin-coregulated pilus (TCP), the genes for which reside in a larger
genetic element, the TCP pathogenicity island. We analyzed 146 CTX-negative strains of V. cholerae O1 or non-O1 isolated
from patients or surface waters in five different countries for the
presence of the TCP pathogenicity island, the regulatory gene
toxR, and the CTX
attachment sequence attRS,
as well as for susceptibility of the strains to CTX
, to investigate
the molecular basis for the emergence of new clones of toxigenic
V. cholerae. DNA probe or PCR assays for tcpA,
tcpI, acfB, toxR, and
attRS revealed that 6.85% of the strains, all of which
belonged to the O1 serogroup, carried the TCP pathogenicity island,
toxR, and multiple copies of attRS, whereas the
remaining 93.15% of the strains were negative for TCP but positive for
either one or both or neither of toxR and
attRS. An analysis of the strains for susceptibility to
CTX
, using a genetically marked derivative of the phage CTX-Km
,
showed that all TCP-positive CTX-negative strains and 1 of 136 TCP-negative strains were infected by the phage either in vitro or in
the intestines of infant mice. The phage genome integrated into the
chromosome of infected V. cholerae O1 cells forming stable
lysogens. Comparative analysis of rRNA gene restriction patterns
revealed that the lysogens derived from nontoxigenic progenitors were
either closely related to or distinctly different from previously
described clones of toxigenic V. cholerae. To our
knowledge, this is the first demonstration of lysogenic conversion of
naturally occurring nontoxigenic V. cholerae strains by
CTX
. The results of this study further indicated that strains belonging to the O1 serogroup of V. cholerae are more
likely to possess the TCP pathogenicity island and hence to be infected by CTX
, leading to the origination of potential new epidemic clones.
*
Corresponding author. Mailing address: Molecular
Genetics Laboratory, Laboratory Sciences Division, ICDDR,B, GPO Box
128, Dhaka-1000, Bangladesh. Phone: 880 2 871751 to 880 2 871760. Fax: 880 2 872529 and 880 2 883116. E-mail: faruque{at}icddrb.org.
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