Detection of toxigenic Vibrio cholerae O1 using polymerase chain reaction for amplifying the cholera enterotoxin gene.
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概要
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A rapid and simple procedure using polymerase chain reaction (PCR) was developed for the detection of cholera enterotoxin (CT) producing character. This method is based on amplifying a 380 base pair (bp) segment of the CT gene (<I>ctx</I>) which controls the production of CT.<BR>Two single-stranded oligonucleotides, synthetized to be complementary to the known nucleotide sequences of genes encoding the A-subunit of ctx, were used as extension primers. The oligonucleotide sequences are 5'TCAAACTATATTGTCTGGTC (CT-1) and 5'CGCAAGTATTACTCATCGA (CT-2). As template DNA was used 5, u1 of boiled bacterial culture broth at 95°C for 5 min without the need for DNA extraction.<BR>The amplified target DNA were confirmed with only CT producing Vibrio cholerae o1 but not with CT non-producing organisms such as heat labile enterotoxin producing Escherichia coli by electrophoretic analysis of PCR mixture after amplification. A few isolates of CT producing V. mimicus and <I>V. cholerae</I> non-O1 were identified.
- The Japanese Association for Infectious Diseasesの論文
The Japanese Association for Infectious Diseases | 論文
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