title: Detection of Vibrio cholerae by isothermal cross-priming amplification combined with nucleic acid detection strip analysis.
abstract: Vibrio cholerae is a water- and food-borne human pathogen, and V. cholerae serotypes O1 and O139 have attracted attention because of their severe pathogenesis. However, non-O1, non-O139 cholera vibrios (NCVs) were also recently recognized as having virulence properties. In this study, we developed a cross-priming amplification (CPA) method for the detection of all serotypes of V. cholerae. The specificity of the CPA method was tested using a panel of 60 different bacterial strains. All of the V. cholerae strains showed positive results, and 41 other types of bacteria gave negative results. The limit of detection of the CPA method was 79.28 fg of genomic DNA, 4.2 × 10(2) CFU/ml for bacteria in pure culture, and 5.6 CFU per 25 g of sample with pre-enrichment. This method showed a higher sensitivity than the loop-mediated isothermal amplification (LAMP) method did and was more convenient to perform. These results indicate that the CPA method can be used for the rapid preliminary screening of V. cholerae.
authors.affiliation: Key Laboratory of Food Nutrition and Safety, Tianjin University of Science and Technology, Tianjin 300457, China; Tianjin Entry-Exit Inspection and Quarantine Bureau, Tianjin 300461, China.
authors.forename: Xia
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Food Nutrition and Safety, Tianjin University of Science and Technology, Tianjin 300457, China.
authors.forename: Xin-Jun
authors.initials: XJ
authors.lastname: Du
authors.affiliation: Tianjin Entry-Exit Inspection and Quarantine Bureau, Tianjin 300461, China.
authors.forename: Chun
authors.initials: C
authors.lastname: Guan
authors.affiliation: Key Laboratory of Food Nutrition and Safety, Tianjin University of Science and Technology, Tianjin 300457, China.
authors.forename: Ping
authors.initials: P
authors.lastname: Li
authors.affiliation: Tianjin Entry-Exit Inspection and Quarantine Bureau, Tianjin 300461, China.
authors.forename: Wen-Jie
authors.initials: WJ
authors.lastname: Zheng
authors.affiliation: Key Laboratory of Food Nutrition and Safety, Tianjin University of Science and Technology, Tianjin 300457, China. Electronic address: s.wang@tust.edu.cn.
authors.forename: Shuo
authors.initials: S
authors.lastname: Wang
mesh_terms: D005516:Food Microbiology; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D009696:Nucleic Acids / Q000032:analysis*; D014734:Vibrio cholerae / Q000235:genetics / Q000302:isolation & purification*; D014871:Water Microbiology
chemical_list: D009696:Nucleic Acids
keywords: Isothermal cross-priming amplification; Strip detection; Vibrio cholerae