Pcr

Multiplex PCR was successfully used to detect the CT and associated virulence gene fragments and, simultaneously,

Fig. 2 Agarose gel electrophoresis of hexaplex PCR products obtained with V. cholerae strains. Lane M, 100-bp DNA ladder (NEB); lanes 1 and 2, toxigenic V. cholerae O1 El Tor strain 20 and classical strain 569B; lanes 3-5, nontoxigenic V. cholerae O1 El Tor strains X392, 2740-80, and 1074-78; lane 6, tcpA~ V. cholerae O1 classical strain O395 RT 110-12; lane 7, toxR~ V. cholerae O1 classical strain O395-12; lane 8, ctxA~V. cholerae O1 classical strain CVD 103-HgR.

Fig. 2 Agarose gel electrophoresis of hexaplex PCR products obtained with V. cholerae strains. Lane M, 100-bp DNA ladder (NEB); lanes 1 and 2, toxigenic V. cholerae O1 El Tor strain 20 and classical strain 569B; lanes 3-5, nontoxigenic V. cholerae O1 El Tor strains X392, 2740-80, and 1074-78; lane 6, tcpA~ V. cholerae O1 classical strain O395 RT 110-12; lane 7, toxR~ V. cholerae O1 classical strain O395-12; lane 8, ctxA~V. cholerae O1 classical strain CVD 103-HgR.

to identify the O1 and O139 serogroups of V. cholerae.[20] From the diagnostic point of view, toxigenic-pathogenic and nontoxigenic-nonpathogenic strains of V. cholerae can be differentiated by the presence of ctx and tcp genes. It was difficult to know whether the CT gene-negative strain possesses other toxin genes, such as, zot, ace, part of the CTX genetic element, and ompU. None of these methods can be used to individually identify toxin and virulence genes. Hence, a one-step hexaplex PCR was developed for rapid detection of an array of six specific virulence genes comprising CT subunit A (ctxA), zonula occludens toxin (zot), accessory cholera enterotoxin (ace), adherence outer membrane protein (ompU), TCP (tcpA), and the regulatory protein (ToxR).[20] Results of representative strains of V. cholerae are shown in Fig. 2. Employing the hexaplex PCR, 97 strains of V. cholerae and Vibrio mimicus were screened for the presence of virulence genes. It was found that hexaplex PCR can rapidly detect, in a single tube, the six commonly known virulence genes in V. cholerae, and was specific and sensitive.

Getting Started With Dumbbells

Getting Started With Dumbbells

The use of dumbbells gives you a much more comprehensive strengthening effect because the workout engages your stabilizer muscles, in addition to the muscle you may be pin-pointing. Without all of the belts and artificial stabilizers of a machine, you also engage your core muscles, which are your body's natural stabilizers.

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