Rapid visualized isothermal nucleic acid testing of Vibrio parahaemolyticus by polymerase spiral reaction
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The aim of this study was to develop an effective and specific visual method to rapidly detect and identify Vibrio parahaemolyticus (V. parahaemolyticus) based on the polymerase spiral reaction (PSR). The method utilized only two pairs of primers designed specifically to target the conserved tlh gene sequence of V. parahaemolyticus. Nucleic acid amplification can be achieved under isothermal conditions using DNA polymerase. The reaction could be accomplished in < 40 min with high specificity and sensitivity. The limits of detection of V. parahaemolyticus in purified genomic DNA and pure culture were 300 fg/μL and 2.4 CFU/mL per reaction, respectively, which were 100-fold more sensitive than with conventional PCR. The model food samples showed consistent specificity and sensitivity to the pure bacterial culture. With these encouraging results, it is expected that the novel, effortless and reliable isothermal nucleic acid testing assay developed in this study has potential to be applied to screening for V. parahaemolyticus in seafood samples.
KeywordsVibrio parahaemolyticus Polymerase spiral reaction Isothermal nucleic acid testing Rapid detection
This study was funded by the National Natural Science Foundation of China (Grant numbers 81502849 and 81872668), the Bethune Medical Scientific Research Fund Project of Jilin University (Grant number 2018B20), the Scientific and Technological Research Project of Jilin Province (Grant numbers 20170204003SF and 20180101095JC), Health science and technology capacity improvement project of Jilin Province (2019Q011) and the Fundamental Research Funds for the Central Universities.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
- 1.Cruz FR, Mai HN, Dhar AK. Multiplex SYBR Green and duplex TaqMan real-time PCR assays for the detection of Photorhabdus insect-related (Pir) toxin genes pirA and pirB. Mol Cell Probes. 2019;43:20–8.Google Scholar
- 3.Aguirre GG, Vazquez JR, Ascencio F. Differences in the susceptibility of American white shrimp larval substages (Litopenaeus vannamei) to four Vibrio species. J Invertebr Pathol. 2001;78:215–9.Google Scholar
- 12.UNICEF/UNDP/World Bank/WHO special program for research and training in tropical diseases. World Health Organization.Google Scholar
- 13.Marvin D, Derong Z, Dayang L, Ningwei H, Xiaoming AA, Big Y, et al. Polymerase reaction spiral rapid detection of influenza A (H1N1) virus. Mil Med. 2017;41:449–52.Google Scholar
- 20.Wang S, Yang J, Shen Z, et al. Analysis of 766 cases of bacterial food poisoning in China from 1994 to 2003. China Prev Med. 2006;180–184.Google Scholar
- 21.Yan C, Yunchang G, Zhutian W, Xm L, Hong L, Month WA, et al. Analysis of surveillance data on foodborne disease outbreaks in China in 2006. Health Res. 2010;39:331–4.Google Scholar