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海产鱼类中异尖线虫酶消化检测技术的研究与应用
引用本文:许旭,黄维义,隋建新,林洪,鲜均章,励建荣,曹立民.海产鱼类中异尖线虫酶消化检测技术的研究与应用[J].中国海洋大学学报(自然科学版),2010,40(3).
作者姓名:许旭  黄维义  隋建新  林洪  鲜均章  励建荣  曹立民
作者单位:1. 中国海洋大学水产品安全实验室,山东,青岛,266003
2. 广西大学动物科学学院,广西,南宁,530004
3. 浙江工商大学食品与生物工程学院,浙江,杭州,310035
基金项目:国家高技术研究发展计划项目,国家鲆鲽类产业技术体系项目 
摘    要:以3种海鱼(牙鲆、狭鳕、鲱鱼)为样本,针对海产品中主要的致病性寄生虫-异尖线虫,开展了酶消化检测技术的研究.根据鱼肉消化前后干物质的质量变化设计了胃蛋白酶消化效率的计算方法,并按照鱼肉:消化液=1:10 (g/mL)的反应比例,分别确定了酶水解鱼肉的最佳条件为:初始pH值为1.1,温度为37 ℃,酶活力为8 U/mL左右;消化后所得的虫体采用多重PCR方法替代传统的形态学观察进行种属鉴定,从而初步建立了基于酶水解-多重PCR的异尖线虫的酶消化检测体系.实际样本检测表明,该技术可以较为准确、快速地用于海产鱼类中简单异尖线虫(Anisakis simplex)和伪地新线虫(Pseudoterranova decipiens)的确证性检测.

关 键 词:异尖线虫  海产鱼类  检测  酶消化  多重PCR

Detection of Anisakid Larvae in Fishes by Enzymatic Degradation
XU Xu,HUANG Wei-Yi,SUI Jian-Xin,LIN Hong,XIAN Jun-Zhang,LI Jian-Rong,CAO Li-Min.Detection of Anisakid Larvae in Fishes by Enzymatic Degradation[J].Periodical of Ocean University of China,2010,40(3).
Authors:XU Xu  HUANG Wei-Yi  SUI Jian-Xin  LIN Hong  XIAN Jun-Zhang  LI Jian-Rong  CAO Li-Min
Institution:XU+Xu1,HUANG+Wei-Yi2,SUI+Jian-Xin1,LIN+Hong1,XIAN+Jun-Zhang1,LI+Jian-Rong3,CAO+Li-Min1(1.Food+Safety+Laboratory,Ocean+University+of+China,Qingdao+266003,China,2.School+of+Animal+Science,Guangxi+University,Nanning+530004,3.School+of+Food+Science+,+Biotechnology,Zhejiang+Gongshang+University,Hangzhou+310035,China)
Abstract:In this study an enzymatic degradation-based method was developed for confirmatory analysis of anisakid larvae, one of the important pathogenic parasites in sea foods. The digestive efficiency was calculated based on the weight changes of fish muscles during enzymatic degradation. When the ratio of fillets to digestive solutions was 1:10 (g/mL), the optimal initial pH was found to be 1.1, the optimal temperature was about 37 ℃ and the enzyme activity should be controlled at 8 U/mL to reach the best digestive effect. After enzymatic degradation, the were collected and then identified by multiplex PCR instead of traditional morphological methods, which was then combined with enzymatic process to construct a new analytical system for the detection of anisakid larvae in sea foods. The established method was preliminarily validated with real fish samples, and proved to be suitable for confirmatory analysis of Anisakis simplex and Pseudoterranova decipiens in these fishery products.
Keywords:anisakid larvae  fish  detection  enzymatic degradation  multiplex PCR
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