Development of a rapid isothermal amplification method for clinical detection of Vibrio cholerae.

INTRODUCTION: The aim of this study was to develop a rapid isothermal amplification assay for Vibrio cholerae. METHODOLOGY: Specific primers were designed following the guidelines of the National Center for Biotechnology Information (NCBI) database and based on the highly conserved sequence of the Vibrio cholerae virulence factor gene ctxA. A detection method was then developed using recombinase polymerase amplification (RPA). Sensitivity, specificity, clinical sample collection, and processing were calculated. RESULTS: A rapid isothermal amplification system was developed, showing a detection capability of one copy per microliter. Specificity analysis against a panel of control strains yielded no false-positive results under the tested conditions. The presence of V. cholerae was detected in 9 fecal samples collected from 20 patients who were hospitalized with suspected cholera infection. CONCLUSIONS: This method can be an effective aid in the detection of bacteria in clinical samples.