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논문 기본 정보

자료유형
학술저널
저자정보
Hu Yuan-qing (School of Biological Science and Biotechnology Minnan Normal University Zhangzhou 363000 P.R. China) Huang Xian-hui (School of Biological Science and Biotechnology Minnan Normal University Zhangzhou 363000 P.R. China) Guo Li-qing (Zhangzhou Center for Disease Control and Prevention Zhangzhou 363000 P.R. China) Shen Zi-chen (School of Biological Science and Biotechnology Minnan Normal University Zhangzhou 363000 P.R. China) LV Lin-xue (School of Biological Science and Biotechnology Minnan Normal University Zhangzhou 363000 P.R. China) Li Feng-xia (School of Biological Science and Biotechnology Minnan Normal University Zhangzhou 363000 P.R. China) Zhou Zan-hu (Comprehensive Technical Service Center Zhangzhou Customs Zhangzhou 363000 P.R. China) Zhang Dan-feng (School of Biological Science and Biotechnology Minnan Normal University Zhangzhou 363000 P.R. China)
저널정보
한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제31권 제12호
발행연도
2021.12
수록면
1,672 - 1,683 (12page)
DOI
10.4014/jmb.2107.07022

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Vibrio parahaemolyticus is recognized as one of the most important foodborne pathogens responsible for gastroenteritis in humans. The blaCARB-17 gene is an intrinsic β-lactamase gene and a novel species-specific genetic marker of V. parahaemolyticus. In this study, a loop-mediated isothermal amplification (LAMP) assay combined with a lateral flow dipstick (LFD) was developed targeting this blaCARB-17 gene. The specificity of LAMP-LFD was ascertained by detecting V. parahaemolyticus ATCC 17802 and seven other non-V. parahaemolyticus strains. Finally, the practicability of LAMP-LFD was confirmed by detection with V. parahaemolyticus-contaminated samples and natural food samples. The results showed that the optimized reaction parameters of LAMP are as follows: 2.4 mmol/l Mg2+, 0.96 mmol/l dNTPs, 4.8 U Bst DNA polymerase, and an 8:1 ratio of inner primer to outer primer, at 63oC for 40 min. The optimized reaction time of the LFD assay is 60 min. Cross-reactivity analysis with the seven non-V. parahaemolyticus strains showed that LAMP-LFD was exclusively specific for V. parahaemolyticus. The detection limit of LAMP-LFD for V. parahaemolyticus genomic DNA was 2.1 × 10-4 ng/μl, corresponding to 630 fg/reaction and displaying a sensitivity that is 100-fold higher than that of conventional PCR. LAMP-LFD in a spiking study revealed a detection limit of approximately 6 CFU/ml, which was similar with conventional PCR. The developed LAMP-LFD specifically identified the 10 V. parahaemolyticus isolates from 30 seafood samples, suggesting that this LAMP-LFD may be a suitable diagnostic method for detecting V. parahaemolyticus in aquatic foods.

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