贝类单孢子虫PCR检测方法的建立

›› 2010, Vol. 37 ›› Issue (1): 54-56.

贝类单孢子虫PCR检测方法的建立

谢丽基，谢芝勋，庞耀珊，刘加波，邓显文，谢志勤

（广西兽医研究所，南宁 530001）

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2010-01-20 发布日期:2010-01-20
通讯作者: 谢芝勋

Development of A Polymerase Chain Reaction Assay for Detection of Haplosporidium in Shellfish

XIE Li-ji, XIE Zhi-xun, PANG Yao-shan, LIU Jia-bo, DENG Xian-wen, XIE Zhi-qin

(Guangxi Veterinary Research Institute, Nanning 530001, China)

Received:1900-01-01 Revised:1900-01-01 Online:2010-01-20 Published:2010-01-20
Contact: XIE Zhi-qin

摘要/Abstract

摘要： 根据基因库中单孢子虫的基因保守序列，设计了1对特异性引物，通过对PCR扩增条件的优化，建立了检测贝类单孢子虫的PCR方法。用该方法对单孢子虫模板进行扩增，得到与试验设计相符的244 bp的特异性条带，而对派琴虫、折光马尔太虫、嗜水气单胞菌、荧光假单胞菌、副溶血弧菌、溶藻弧菌、河弧菌和拟态弧菌等病原体的扩增结果全为阴性。敏感性试验结果表明，该方法最低能检测到100 fg的单孢子虫DNA。

关键词: 贝类; 单孢子虫; PCR检测方法

Abstract: According to the gene sequences in GenBank of Haplosporidium sp, a pair of specific primers were designed for amplifying the specific fragments of Haplosporidium sp. The reaction parameters were optimized to develop the polymerase chain reaction method for detection of Haplosporidium sp. The 244 bp long specific DNA fragment of Haplosporidium sp was amplified, but not from other pathogenic such as Perkinsus sp, Marteilia refringens, Aeromonas hydrophila, Pseudomonas fluorescens, Vibrio parahaemolyticu, Vibrio Alginolyticu, Vibrio Fluvialis and Vibrio Mimicus by PCR. The sensitivity results showed that as little as 100 fg DNA of Haplosporidium sp was detected by this PCR.

Key words: Shellfish; Haplosporidium sp; PCR

中图分类号:

S852.7

谢丽基;谢芝勋;庞耀珊;刘加波;邓显文;谢志勤. 贝类单孢子虫PCR检测方法的建立[J]. , 2010, 37(1): 54-56.

XIE Li-ji;XIE Zhi-xun;PANG Yao-shan;LIU Jia-bo;DENG Xian-wen;XIE Zhi-qin. Development of A Polymerase Chain Reaction Assay for Detection of Haplosporidium in Shellfish[J]. China Animal Husbandry & Veterinary Medicine, 2010, 37(1): 54-56.

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