References in periodicals archive ?
In the present study, the IgY technology has been employed for the production of specific antibodies against OmpW and CtxB antigens of V.
One colony grown on BHI plates was suspended in 200 [micro]L of TE 10: 1 (10 mM Tris-HCl pH 8.0, 1mM EDTA), boiled for 20 minutes, and immediately used in PCR reactions with primers designed for the amplification of ctxA and tcpA 13], zot and ace , ctxB 15], and rfbN 16].
Based on Vibrio phage CTX strain IB1482, specific primers were designed to amplify coding sequence of ctxB. The expected size of PCR product, approximately 330 bp, was obtained following PCR (Figure 1A).
cholerae strains were examined for the presence of ctxA, ctxB, zot, ace, toxR, [tcpA.sup.C], [tcpA.sup.E], [hlyA.sup.C] and [hlyA.sup.E] by conventional PCR using strains AR15493 and AR15425 from Bangladesh as positive controls for zot, ace, toxR, and hlyA genes and strain C6706 as positive control for ctxAB and tcpA (19).
We studied the genotype of ctxB using a double-mismatch amplification mutation PCR (i.e., mismatches in both primers).
Classical ctxB in Vibrio cholerae O1, Kolkata, India.
The ctxB genotyping of the selected strains was performed by mismatch amplification mutation (MAMA)-PCR assay (33).
cholerae O1 strain PCR tcp tcp Strain ompW toxR [A.sup.CL] [A.sup.EI] Env-9 * + + + - Env-90 + + - + Env-94 + + - + Env-122 *([dagger]) + + + - Env-383 + + - + Env-390 * + + + - Env-114 *([dagger]) + + + - PCR rst Strain ctxA ctxB [R.sup.I] Env-9 * - - - Env-90 + + + Env-94 + + + Env-122 *([dagger]) - - - Env-383 + + + Env-390 * - - - Env-114 *([dagger]) - - - PCR rst rst rst Strain [R.sup.CL] [C.sup.EI] [C.sup.CL] Env-9 * - - - Env-90 - - - Env-94 - - - Env-122 *([dagger]) - - - Env-383 - - - Env-390 * - - - Env-114 *([dagger]) - - - Mismatch amplification mutation assay PCR ctx ctx Strain [B.sup.CL] [B.sup.EI] Env-9 * - - Env-90 + - Env-94 + - Env-122 *([dagger]) - - Env-383 + - Env-390 * - - Env-114 *([dagger]) - - * Indicates ctx-negative V.
Another classification identifies three types of ctxB genes based on three non random base changes resulting in modifications in the deduced amino acid sequence.
Historically, CTX prophages have been categorized as [CTX.sup.classical] or [CTX.sup.El Tor] on the basis of DNA sequence of the rstR and the sequence variation in ctxB gene.
cholerae ET has shown genetic changes since 2001, and isolates carry the ctxB gene of the CL biotype ([ctxB.sup.CL]) in Bangladesh (4).
To determine the genetic relatedness of the isolates, pulsed-field gel electrophoresis (PFGE) was performed according to the established PulseNet protocol (9) and analyzed with BioNumerics 6.0 (Applied Maths, Kortrijk, Belgium); ctxB genotyping was also performed as described (10).
Acronyms browser ?
Full browser ?