Evaluation of a novel lysis-based sample processing method to optimize Vibrio vulnificus detecting by Loop-Mediated Isothermal Amplification assay

Abstract

Abstract Purpose To explore a rapid sample preprocessing method suitable for Vibrio vulnificus detecting by Loop-Mediated Isothermal Amplification assay to shorten the total detecting time and increase efficiency. Methods A water-lysis sample processing method was established by optimizing the bacteria/water ratio, and V. vulnificus, the specific pathogen, was detected using the lysis-based LAMP assay established in our study. To evaluate its efficiency, pure bacteria gradient dilution and commercial products were detected for specificity and sensitivity. V. parahemolyticus, V .alginolyticus, and Proteus mirabilis were used to confirm its broad-spectrum application. Result The water-lysis-based V. vulnificus detecting LAMP method shortened preprocessing time to ≤1 min with 100% LAMP specificity; the detection limits of the LAMP assay were decreased to 1.2 × 102 CFU/mL in pure culture, and 1.47 × 103 CFU/g in oyster, respectively. Furthermore, the 100% LAMP specificity and high sensitivity of the water-lysis method were also obtained on detecting V. parahaemolyticus, V. alginolyticus, and P. mirabilis, revealing its excellent LAMP adaption with improvement in sensitivity and efficiency. Conclusion Our study provided a novel LAMP preprocessing method that was faster and more precise compared to common methods and possessed the practical potential for LAMP application in the future.