Detection of Vibrio cholerae by isothermal cross-priming amplification combined with nucleic acid detection strip analysis

Xia Zhang; Xin-Jun Du; Chun Guan; Ping Li; Wen-Jie Zheng; Shuo Wang

doi:10.1016/j.mcp.2015.05.001

Detection of Vibrio cholerae by isothermal cross-priming amplification combined with nucleic acid detection strip analysis

Mol Cell Probes. 2015 Aug;29(4):208-14. doi: 10.1016/j.mcp.2015.05.001. Epub 2015 May 8.

Authors

Xia Zhang¹, Xin-Jun Du², Chun Guan³, Ping Li², Wen-Jie Zheng³, Shuo Wang⁴

Affiliations

¹ 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.
² Key Laboratory of Food Nutrition and Safety, Tianjin University of Science and Technology, Tianjin 300457, China.
³ Tianjin Entry-Exit Inspection and Quarantine Bureau, Tianjin 300461, China.
⁴ Key Laboratory of Food Nutrition and Safety, Tianjin University of Science and Technology, Tianjin 300457, China. Electronic address: s.wang@tust.edu.cn.

PMID: 25962828
DOI: 10.1016/j.mcp.2015.05.001

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.

Keywords: Isothermal cross-priming amplification; Strip detection; Vibrio cholerae.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Food Microbiology
Nucleic Acid Amplification Techniques / methods*
Nucleic Acids / analysis*
Vibrio cholerae / genetics
Vibrio cholerae / isolation & purification*
Water Microbiology

Substances

Nucleic Acids