Quantitative detection method of Enterocytozoon hepatopenaei using TaqMan probe real-time PCR

文献类型： 外文期刊

第一作者： Liu, Ya-Mei

作者： Liu, Ya-Mei;Qiu, Liang;Sheng, An-Zhi;Wan, Xiao-Yuan;Cheng, Dong-Yuan;Huang, Jie;Liu, Ya-Mei;Qiu, Liang;Cheng, Dong-Yuan;Huang, Jie

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关键词： TaqMan-based qPCR;Enterocytozoon hepatopenaei (EHP);Body mass index (BMI)

期刊名称：JOURNAL OF INVERTEBRATE PATHOLOGY （ 影响因子：2.841； 五年影响因子：3.368 ）

ISSN： 0022-2011

年卷期： 2018 年 151 卷

页码：

收录情况： SCI

摘要： A TaqMan probe and a pair of specific primers were selected from the small subunit ribosomal DNA (SSU rDNA) sequence of Enterocytozoon hepatopenaei (EHP); this real-time PCR assay was developed and optimized. It showed a good linearity in detecting standards of EHP SSU rDNA fragments from 4 x 10(2) to 4 x 10(8) copies/reaction using the established method. The detection limit of the qPCR method was as low as 4 x 10(1) copies per reaction, which was higher than the conventional PCR and SYBR Green I-based EHP qPCR reported. Using the qPCR assay, EHP was detected in four batches of slow-growing Penaeus vannamei specimens collected from Tianjin and Zhejiang Province in China was detected using qPCR. The results showed that all the hepatopancreas from the slow-growing P. vannamei specimens were detected as EHP-positive. EHP copies of hepatopancreas in some batches had a negative correlation with the body mass index (BMI) of shrimps; however, not all batches of specimens had this negative correlation between EHP copies of hepatopancreas and BMI. This qPCR technique is sensitive, specific and easy to perform (96 tests in < 3 h), which provides technical support for the detection and prevention of EHP.

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