Rapid and simultaneous detection of virulence factors of Vibrio parahaemolyticus by multiplex microfluidic-LAMP (MMLAMP)

被引：7

作者：

Lang, Ziyue ^{[1
]}

Ye, Xin ^{[2
]}

Liu, Jia ^{[1
]}

Lin, Shoujia ^{[1
]}

Li, Yunfei ^{[3
]}

Chen, Qi ^{[1
]}

Cao, Hongmei ^{[1
]}

机构：

[1] Hainan Univ, Coll Food Sci & Engn, Haikou 570228, Peoples R China

[2] Xi An Jiao Tong Univ, Dept Lab Med, Affiliated Hosp 1, Xian 710061, Peoples R China

[3] Hainan Univ, Coll Biomed Engn, Haikou 570228, Peoples R China

来源：

MICROCHEMICAL JOURNAL | 2024年 / 201卷

基金：

中国国家自然科学基金;

关键词：

Vibrio parahaemolyticus; Microfluidic chip; Loop-mediated isothermal amplification; Virulence factor; MEDIATED ISOTHERMAL AMPLIFICATION; IMMUNOMAGNETIC SEPARATION; PCR; GENE; CHIP;

D O I：

10.1016/j.microc.2024.110738

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

Vibrio parahaemolyticus (VP) is an essential foodborne bacterium who can cause gastroenteritis. However, current detection methods generally still have the difficulty for quickly screening VP strains and accurately detecting VP. In this work, a sensitive and specific multiplex microfluidic-LAMP (MMLAMP) method was constructed to rapidly detect VP using a centrifugal microfluidic chip combined with LAMP. The system enables simultaneous detection of two hemolysin genes (tlh and trh) and one VP regulatory factor (toxR) in a single chip improving the accuracy for detecting VP. The MMLAMP system detected 2.42 x 10(4) copies/mu L of the plasmid (tlh-pUC57), 4.44 x 10(3) copies/mu L of the plasmid (trh-pUC57) and 4.86 x 10(4) copies/mu L of the plasmid (toxR-pUC57) in less than 35 min, respectively. Meanwhile, MMLAMP system was successfully applied for VP detection in artificially contaminated samples, the LOD is 220 CFU/g, which is 10 times lower than that of qPCR within the same detection time. The system also demonstrated 100 % specificity and good precision (C.V. < 5 %) in detecting the three targets of the VP, providing a new approach for point-of-care testing (POCT) of VP.

引用

页数：8

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