A simple PCR method for the detection of pathogenic spiroplasmas in crustaceans and environmental samples
A simple PCR method for the detection of pathogenic spiroplasmas in crustaceans and environmental samples
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DOI：
10.1016/j.aquaculture.2007.01.032
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发表时间：
2007-05-01
期刊：
影响因子：
4.5
通讯作者：
Chen, Jianxiu
中科院分区：
文献类型：
--
作者：
Ding, Zhengfeng;Bi, Keran;Chen, Jianxiu
一个简单的PCR方法，使用Chelex-100提取已被开发用于检测在商业上重要的甲壳类动物中的致病性螺原体的16 STRNA基因。该方法检测了中华绒螯蟹中华绒螯蟹，克氏原螯虾，以及从宿主栖息地的环境样品中的螺原体。与组织/细胞基因组DNA分离试剂盒法相比，Chelex-100法对所有来源的螺原体的检测更敏感，可在4 h内获得结果。检测限为1 CCU/mL。这是首次使用Chelex-100提取方法检测养殖甲壳动物中的螺原体。这是一种简单、快速、灵敏和低成本的方法，可用于广泛的实地调查。(c)2007 Elsevier B. V.保留所有权利。
A simple PCR method that uses a Chelex-100 extraction has been developed to detect the 16STRNA gene of pathogenic spiroplasmas in commercially important crustaceans. The method detected spiroplasmas in the Chinese mitten crab Eriocheir sinensis, and the crayfish Procambarus clarki, as well as from environmental samples from host habitats. Compared with the Tissues/Cell Genomic DNA Isolation Kit method, the Chelex-100 method was more sensitive to the detection of spiroplasmas from all sources sampled, with results being obtained within 4 h. The detection limit was 1 CCU/mL. This is the first time that the Chelex-100 extraction method has been used to detect spiroplasmas from cultured crustaceans. It is a simple, rapid, sensitive and low-cost method and can be applied to broad, field-based surveys. (c) 2007 Elsevier B.V. All rights reserved.