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当前位置: 首页 > 文章 > 贝类单孢子虫二温式PCR检测方法的建立及初步应用 黑龙江畜牧兽医 2013,49 (13) 73-75
Position: Home > Articles > Establishment and primary application of a two-temperature polymerase chain reaction assay for detection of Haplosporidium sp. in shellfish Heilongjiang Animal Science and Veterinary Medicine 2013,49 (13) 73-75

贝类单孢子虫二温式PCR检测方法的建立及初步应用

作  者：
谢丽基;谢芝勋;刘加波;庞耀珊;邓显文;谢志勤;范晴
单  位：
广西壮族自治区兽医研究所
关键词：
贝类;单孢子虫;二温式PCR
摘  要：
为了建立检测贝类单孢子虫的二温式PCR方法,试验根据GenBank中单孢子虫的基因保守序列设计了1对特异性引物,并对二温式PCR扩增条件进行优化。结果表明:用二温式PCR方法对单孢子虫模板进行扩增,得到与试验设计相符的333 bp特异性扩增带,而对折光马尔太虫、派琴虫、副溶血弧菌、溶藻弧菌和河弧菌等病原体进行扩增,结果全为阴性;该方法最低能检测到1 fg的单孢子虫质粒DNA。
译  名：
Establishment and primary application of a two-temperature polymerase chain reaction assay for detection of Haplosporidium sp. in shellfish
作  者：
XIE Li-ji,XIE Zhi-xun,LIU Jia-bo,PANG Yao-shan,DENG Xian-wen,XIE Zhi-qin,FAN Qing(Guangxi Key Laboratory of Animal Vaccines and New Technology,Guangxi Veterinary Research Institute,Nanning 530001,China)
关键词：
shellfish;Haplosporidium sp.;two-temperature PCR
摘  要：
To establish the two-temperature PCR method for detection of Haplosporidium sp.,a pair of specific primers was designed according to the conserved gene sequence of Haplosporidium sp.published in GenBank,and then the amplification conditions of the two-temperature PCR method were optimized.The results showed that the specific DNA fragment of Haplosporidium sp.with the length of 333 bp was obtained using the PCR method for the amplification of Haplosporidium sp.as a template,which was consistent with the design of the test,but other pathogens such as Marteilia refringens,Perkinsus sp.,Vibrio parahaemolyticus,Vibrio aginolyticus,and Vibrio fluvialis were found to have negative results in the amplification by this two-temperature PCR.The results indicate that the two-temperature PCR method has the lowest sensitivity;it can detect 1fg of plasmid DNA of Haplosporidium sp.

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