Development of a highly sensitive lateral immunochromatographic assay for rapid detection of Vibrio parahaemolyticus

• Published in: Journal of microbiological methods Vol. 131; pp. 78 - 84
• Main Authors: Liu, Xinfeng, Guan, Yuyao, Cheng, Shiliang, Huang, Yidan, Yan, Qin, Zhang, Jun, Huang, Guanjun, Zheng, Jian, Liu, Tianqiang
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.12.2016
• Subjects: Animals; Antibodies, Monoclonal; Aquatic Organisms - microbiology; Bacterial Typing Techniques - methods; Cell Line; Decapoda; Diarrhea - microbiology; Disease Models, Animal; Feces - microbiology; Female; Food Safety; Gold Colloid - chemistry; Humans; Hybridomas - microbiology; Immunochromatographic assay; Immunochromatography - instrumentation; Immunochromatography - methods; Immunochromatography - veterinary; Lateral flow device; Mice, Inbred BALB C; Monoclonal antibody; Palaemonidae; Penaeidae - microbiology; Rapid diagnosis; Seafood - microbiology; Sensitivity and Specificity; Species Specificity; Time Factors; Vibrio; Vibrio Infections - diagnosis; Vibrio Infections - immunology; Vibrio Infections - microbiology; Vibrio Infections - veterinary; Vibrio parahaemolyticus; Vibrio parahaemolyticus - classification; Vibrio parahaemolyticus - isolation & purification; Vibrio parahaemolyticus - pathogenicity; Rapid diagnosis; Vibrio parahaemolyticus; Monoclonal antibody; Immunochromatographic assay; Lateral flow device
• ISSN: 0167-7012; 1872-8359
• DOI: 10.1016/j.mimet.2016.10.007

Vibrio parahaemolyticus is widely present in brackish water all over the world, causing infections in certain aquatic animals. It is also a foodborne pathogen that causes diarrhea in humans. The aim of this study is to develop an immunochromatographic lateral flow assay (LFA) for rapid detection of V. parahaemolyticus in both aquatic products and human feces of diarrheal patients. Two monoclonal antibody (MAb) pairs, GA1a-IC9 and IC9-KB4c, were developed and proven to be highly specific and sensitive to V. parahaemolyticus. Based on the two MAb pairs, two types of LFA strips were prepared. Their testing limits for V. parahaemolyticus culture were both 1.2×103CFU/ml. The diagnostic sensitivities and specificities were both 100% for the 32 tested microbial species, including 6 Vibrio species. Subsequently, the LFA strips were used to test Whiteleg shrimps and human feces. The type II strip showed a higher diagnostic sensitivity. Its sensitivity and specificity for hepatopancreas and fecal samples from 13 Whiteleg shrimps and fecal samples from 146 human diarrheal patients were all 100%. In conclusion, our homemade type II LFA is a very promising testing device for rapid and convenient detection of V. parahaemolyticus infection not only in aquatic animals, but also in human diarrheal patients. This sensitive immunochromtographic LFA allows rapid detection of V. parahaemolyticus without requirement of culture enrichment. •Development of MAbs against V. parahaemolyticus•Preparation of lateral flow assay strip to detect V. parahaemolyticus•The detection limit of the assay is 103CFU/ml V. parahaemolyticus•Detection of V. parahaemolyticus infection in shrimps without culture enrichment•Diagnosis of V. parahaemolyticus infection in human feces without culture enrichment