<p><b>OBJECTIVE</b>To develop a real-
time SYBR Green
polymerase chain reaction (
PCR) for
detection of
Vibrio cholerae serogroups O1 and O139, and to evaluate its reliability through
detection of
estuary water samples.</p><p><b>
METHODS</b>
O antigen rfb
genes specific for O1 and O139 were used for the design of
PCR primers. The real-
time SYBR Green
PCR system in detecting O1 and O139 specific rfb
genes in one tube was developed, and its
sensitivity,
specificity and reproducibility were evaluated. The
ability of the
real-time PCR in
detection of
estuary water samples was compared with the routine
PCR and
bacteria isolation.</p><p><b>RESULTS</b>The amplification of O1 or O139 specific target
gene could be detected according to the melt curve
temperature of amplicons. No amplification was observed in the templates of other 10 non-cholerae vibrios. When comparing to the
real-time PCR to
bacteria isolation in
detection of 524
estuary water samples, it showed high
sensitivity, plus also positive in
real-time PCR detection among all the samples in which
bacteria of O1 or O139 were isolated.</p><p><b>CONCLUSION</b>The real-
time SYBR Green
PCR could be used as the first step of rapid
environment screen of V. cholerae in
water samples thus might enhance the
efficiency of isolation in
screening of large amount of
water samples.</p>