对虾急性肝胰腺坏死综合征不同检测方法结果分析

Analysis on the Results of Different Detection Methods for AHPND in Shrimps

急性肝胰腺坏死综合征(acute hepatopancreas necrosis disease,AHPND)是一种危害大、新出现的对虾疫病。目前认为该病由一种副溶血性弧菌(Vibrio parahaemolyticus,VP)引起。本试验通过生化鉴定、血清学分型、MALDI-TOF-MS鉴定以及文献建立的PCR方法,在福建省南美白对虾养殖场,对分离于AHPND病虾肝胰腺的VP进行了检测。通过VITEK-2鉴定出81株VP分离株。从中选取9株分别进行MALDI-TOF-MS鉴定、PCR检测和血清学分型,发现这9株分离菌株均与MALDI-TOF-MS数据库中的VP匹配。其中:4株PCR检测为阳性,被鉴定为AHPND VP,其血清型包括O1:KUT和O1:K68;另5株PCR检测为阴性,血清型包括O1:KUT、O3:K6和O1:K68。试验表明:AHPND VP分离株存在不同血清型,可通过MALDI-TOF-MS进行菌种鉴定;MALDI-TOF-MS与PCR结合使用,可以准确、快速鉴定AHPND VP,这有利于开展AHPND的病原学分析、流行病学调查以及诊断和监测。

At present,acute hepatopancreatic necrosis disease(AHPND),a harmful and emerging shrimps disease,was considered to be caused by Vibrio parahaemolyticus(VP).In this study,the VP strains isolated from the hepatopancreas of shrimps infected with AHPND in a white shrimp farm in Fujian Province were tested by biochemical identification,serological typing,MALDI-TOF-MS and the PCR assay published in related literature.A total of 81 VP isolates were confirmed by VITEK-2,from which,9 strains were selected for MALDI-TOF-MS identification,serological typing and PCR assay,which turned out to be compatible with the VP in MALDI-TOF-MS database.Specifically,according to results of PCR assay,4 strains were detected positive and identified as AHPND VP,and their serotypes were O1:KUT and O1:K68;the other 5 strains were negative with the serotypes of O1:KUT,O3:K6 and O1:K68.In conclusion,there were different serotypes in AHPND VP strains that could be identified by MALDI-TOF-MS;the AHPND VP could be identified accurately and rapidly by combination of MALDI-TOF-MS and PCR assay,which contributed to pathogenic analysis,epidemiological investigation,diagnosis and monitoring against the disease.