Development of both multiple PCR and real-time SYBR green PCR for the detection of Vibrio cholerae non-O1/O139 serogroups / 中华流行病学杂志

Objective To develop methodology of both multiple PCR and real-time SYBR green PCR for the detection of Vibrio cholerae (V.cholerae) serogroups non-O1 and non-O139.Methods The outer membrane protein gene (ompW) specific for V.cholerae,as well as O antigen rfb genes specific for both O1 and O139,were used for the design of the PCR primers.Both multiple PCR and real-time SYBR green PCR systems were used to detect both O1 and O139.Specific rfb genes and ompW were developed to evaluate their specificity,limit of detection,reproducibility and consistency.Results We established multiple PCR and real-time SYBR green PCR methods.According to the specific electrophoretic bands (multiple PCR) and the specific melt curve temperature (real-time SYBR green PCR),both methods could specifically detect the non-O1,non-O139 V.cholerae,and to differentiate them from O1,O139 V.cholerae,other five Vibrios and 3 intestinal bacteria.The detection limits were 7 × 104 cfu/ml (multiple PCR) and 7 × 102 cfu/ml (real-time SYBR green PCR),with statistically significant difference seen (P＜0.05).For the reproducibility of real-time SYBR green PCR,the external coefficient variation ranging from 0.22％ to 0.92％ while the internal coefficient variation ranging from 0.27％ to 1.41％.370 strains of non-O1,non-O139 V.cholerae,were detected,with both consistency rates as 100％.Conclusion Both multiple PCR and real-time SYBR green PCR could detect non-O1,non-O139 V.cholerae,rapidly,specifically,and reproducibly,that could all be used for the detection and identification of non-O 1,non-O 139 under different conditions.

A Case of Vibrio cholerae Non-O1/O139 Gastroenteritis / 대한진단검사의학회지

Vibrio cholerae non-O1/O139 strains are the organisms that are biochemically indistinguishable from V. cholerae but do not agglutinate in Vibrio cholerae O1 and O139 antisera. V. cholerae non-O1/O139 strains are associated with gastroenteritis and extraintestinal infections such as bacteremia, peritonitis and wound infections. Gastroenteritis by V. cholerae non-O1/O139 is uncommon in Korea. We isolated V. cholerae non-O1/O139 from a stool specimen of a one year-old female with diarrhea and high fever.

A Case of Vibrio cholerae non-O1/O139 Peritonitis / 대한임상미생물학회지

Vibrio cholerae strain other than O1 and O139 (Vibrio cholerae non-O1/O139) are associated with sporadic diarrhea and have often been reported in association with extraintestinal infections. We report a case of peritonitis by V. cholerae non-O1/O139 in 43-year-old male who was diagnosed cirrhosis. He, was complained of abdominal distension and fever without history of consumption of raw sea food and exposure to sea water. Gram negative bacilli were cultured from his peritoneal fluid and identified as V. cholerae sero group O14.

A Case of Vibrio cholerae non-O1/O139 Peritonitis / 대한임상미생물학회지

Vibrio cholerae strain other than O1 and O139 (Vibrio cholerae non-O1/O139) are associated with sporadic diarrhea and have often been reported in association with extraintestinal infections. We report a case of peritonitis by V. cholerae non-O1/O139 in 43-year-old male who was diagnosed cirrhosis. He, was complained of abdominal distension and fever without history of consumption of raw sea food and exposure to sea water. Gram negative bacilli were cultured from his peritoneal fluid and identified as V. cholerae sero group O14.