Rapid and Sensitive Detection of Vibrio parahaemolyticus and Vibrio vulnificus by Multiple Endonuclease Restriction Real-Time Loop-Mediated Isothermal Amplification Technique

Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry
Yi WangChangyun Ye

Abstract

Vibrio parahaemolyticus and Vibrio vulnificus are two marine seafood-borne pathogens causing severe illnesses in humans and aquatic animals. In this study, a recently developed novel multiple endonuclease restriction real-time loop-mediated isothermal amplification technology (MERT-LAMP) were successfully developed and evaluated for simultaneous detection of V. parahaemolyticus and V. vulnificus strains in only a single reaction. Two MERT-LAMP primer sets were designed to specifically target toxR gene of V. parahaemolyticus and rpoS gene of V. vulnificus. The MERT-LAMP reactions were conducted at 62 °C, and the positive results were produced in as short as 19 min with the genomic DNA templates extracted from the V. parahaemolyticus and V. vulnificus strains. The two target pathogens present in the same sample could be simultaneously detected and correctly differentiated based on distinct fluorescence curves in a real-time format. The sensitivity of MERT-LAMP assay was 250 fg and 125 fg DNA per reaction with genomic templates of V. parahaemolyticus and V. vulnificus strains, which was in conformity with conventional LAMP detection. Compared with PCR-based techniques, the MERT-LAMP technology was 100- and 10-fold more sensitive t...Continue Reading

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Dec 4, 2015·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Yi WangChangyun Ye

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Citations

Jun 5, 2018·Canadian Journal of Microbiology·Deshun XuLiping Chen
Jan 11, 2020·Frontiers in Microbiology·Ben JiaChaochun Wei
Jun 22, 2021·Frontiers in Microbiology·Shihui FanShujun Wang
Jul 4, 2018·Analytical Chemistry·Sheng CaiAndrew D Ellington

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Datasets Mentioned

BETA
L11929
AY187681

Methods Mentioned

BETA
PCR
electrophoresis
Assay

Software Mentioned

MERT

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