Development and Evaluation of a Loop-Mediated Isothermal Amplification Assay Combined with Enrichment Culture for Rapid Detection of Very Low Numbers of Vibrio parahaemolyticus in Seafood Samples

被引：21

作者：

Di, Huiling ^{[1
]}

Ye, Lei ^{[1
]}

Neogi, Sucharit Basu ^{[1
]}

Meng, Hecheng ^{[2
]}

Yan, He ^{[1
]}

Yamasaki, Shinji ^{[1
,2
]}

Shi, Lei ^{[1
]}

机构：

[1] S China Univ Technol, Coll Light Ind & Food Sci, Guangzhou 510640, Guangdong, Peoples R China

[2] Osaka Prefecture Univ, Grad Sch Life & Environm Sci, Osaka 5988531, Japan

来源：

BIOLOGICAL & PHARMACEUTICAL BULLETIN | 2015年 / 38卷 / 01期

关键词：

Vibrio parahaemolyticus; loop-mediated isothermal amplification; rapid detection; enrichment culture; seafood; THERMOSTABLE DIRECT HEMOLYSIN; REAL-TIME PCR; OYSTERS; VULNIFICUS; GENE; PURIFICATION; SHRIMP; JAPAN; TDH; TRH;

D O I：

10.1248/bpb.b14-00582

中图分类号：

R9 [药学];

学科分类号：

1007 ;

摘要：

The aim of this study was to develop and evaluate a rapid and effective method to detect Vibrio parahaemolyticus, a leading pathogen causing seafood-borne gastroenteritis. A newly designed loop-mediated isothermal amplification (LAMP) assay including a short enrichment period was optimized. This assay correctly detected all the target strains (n=61) but none of the non-target strains (n=34). Very low numbers of V. parahaemolyticus (2 colony forming unit (CFU) per gram of seafood) could be detected within 3 h and the minimum time of the whole assay was only 5h. Comparative screening of various seafood samples (n=70) indicated that the LAMP assay is superior to polymerase chain reaction (PCR) and conventional culture methods because it is more rapid and less complex. This highly sensitive LAMP assay can be applicable as the method of choice in large-scale and rapid screening of seafood and environmental samples to detect V. parahaenwlyticus strains.

引用

页码：82 / 87

页数：6

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