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Fig. S1. Ethidium bromide-stained agarose gel electrophoresis of multiplex PCR products of Vibrio cholerae. Lane M, 100 bp DNA ladder. V. cholerae N16961, VO170 and VOP170 positive controls for CTXΦ, CTXΦ and pre-CTXΦ and pre-CTXΦ alone respectively. V. cholerae VO130 negative control for CTXΦ and pre-CTXΦ. V. cholerae VO239 (O139, Allepey, 2002, Clinical), VO254 (O139, Trivandrum, 2002, Clinical) and VO162 (O139, Wardha, 1997, Clinical) were positive for pre-CTXΦ only. V. cholerae VO250 (O139, Trivandrum, 2002, Clinical), VO165 (O139, Wardha, 1997, Clinical), VO240 (O139, Allepey, 2002, Clinical) and VE234 (O1, Varanasi, 1986, Environmental) were positive for both pre-CTXΦ and CTXΦ, and V. cholerae VO270 (O1, Trivandrum, 2002, Clinical) and VE86 (O1, Varanasi, 1986, Environmental) were positive for CTXΦ.

Fig. S2. Southern blot hybridization of AvaI digested genomic DNA of Vibrio cholerae strains (as in Fig. 1) using (A) ctxA and (B) zot probes. Lane M, HindIII digest of λ DNA. Strains that amplified the 820 bp amplicon in the multiplex PCR showed a single band with ctxA, while those strains that produced only the 115 bp amlicon did not produce any band with ctxA but showed multiple bands when hybridized with zot probe, confirming the presence or absence of pre-CTXΦ. V. cholerae N16961, VO170 and VOP170 positive controls for CTXΦ, CTXΦ and pre-CTXΦ, and pre-CTXΦ alone respectively. V. cholerae VO130 negative control for CTXΦ and pre-CTXΦ. V. cholerae VO239 (O139, Allepey, 2002, Clinical), VO254 (O139, Trivandrum, 2002, Clinical) and VO162 (O139, Wardha, 1997, Clinical) were positive for pre-CTXΦ only. V. cholerae VO250 (O139, Trivandrum, 2002, Clinical), VO165 (O139, Wardha, 1997, Clinical), VO240 (O139, Allepey, 2002, Clinical) and VE234 (O1, Varanasi, 1986, Environmental) were positive for both pre-CTXΦ and CTXΦ, and V. cholerae VO270 (O1, Trivandrum, 2002, Clinical) and VE86 (O1, Varanasi, 1986, Environmental) were positive for CTXΦ.

Table S1. Arrangement of CTXΦ and pre-CTXΦ of V. cholerae O1 and O139 strains used to study MLST, ribotyping, orfU and zot sequence comparison and construction of phylogenetic trees.

Table S2. RFLP patterns obtained using genomic DNA of V. cholerae digested with restriction enzymes and hybridization with ctxA, zot, RS, rstRClass, rstRET, rstRCal respectively.

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Please note: Wiley Blackwell is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.