An improved method for detecting faecal Vibrio cholerae by PCR of the toxin A gene.
A method for removing inhibitor(s) of the PCR assay for the direct detection of cholera toxin A gene (ctxA) in human faeces is described. Inhibitors of the PCR were removed by centrifugation and the activity of the remaining inhibitors by dilution. Based on these data, a protocol was developed for pre-treatment of stool specimens for PCR assay, and a simple and rapid protocol was constructed for the diagnostic detection of the ctxA genes in stool specimens in combination with single band detection on gel electrophoresis, dot-blot hybridisation and enrichment culture. This protocol was applied to clinical specimens and showed that the PCR method gave 100% agreement with established culture methods for the detection of cholera toxin-producing Vibrio cholerae O1. This protocol was considered to be useful because of its simplicity and the rapidity of diagnosis.
Miyagi
Kazufumi
K
Osaka Quarantine Station, 552-0021 Osaka.|Department of Microbiology, Osaka Medical College, 569-8686 Osaka.
Sano
Kouichi
K
Department of Microbiology, Osaka Medical College, 569-8686 Osaka.
Morita
Chizuko
C
Department of Microbiology, Osaka Medical College, 569-8686 Osaka.
Imura
Shunro
S
Kansai (formerly Osaka) Airport Quarantine Station, 549-0011 Osaka.|Department of Microbiology, Osaka Medical College, 569-8686 Osaka.
Morimatsu
Shinichi
S
Department of Microbiology, Osaka Medical College, 569-8686 Osaka.
Goto
Toshiyuki
T
Department of Microbiology, Osaka Medical College, 569-8686 Osaka.
Nakano
Yasuo
Y
Osaka Quarantine Station, 552-0021 Osaka.
Omura
Kanzo
K
Kansai (formerly Osaka) Airport Quarantine Station, 549-0011 Osaka.
Matsumoto
Yasukazu
Y
Kansai (formerly Osaka) Airport Quarantine Station, 549-0011 Osaka.
Maeda
Kenji
K
Kansai (formerly Osaka) Airport Quarantine Station, 549-0011 Osaka.
Hashimoto
Satoru
S
Kansai (formerly Osaka) Airport Quarantine Station, 549-0011 Osaka.
Honda
Takeshi
T
Research Institute for Microbial Diseases, Osaka University, 565-0871 Osaka, Japan.|Department of Microbiology, Osaka Medical College, 569-8686 Osaka.
D002353:Carrier State / Q000382:microbiology; D002771:Cholera / Q000175:diagnosis* / Q000382:microbiology; D002772:Cholera Toxin / Q000235:genetics*; D004587:Electrophoresis, Agar Gel; D005243:Feces / Q000382:microbiology*; D006801:Humans; D007841:Latex Fixation Tests; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014734:Vibrio cholerae / Q000235:genetics / Q000302:isolation & purification*
D002772:Cholera Toxin