Journal
JOURNAL OF INVERTEBRATE PATHOLOGY
Volume 151, Issue -, Pages 191-196Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.jip.2017.12.006
Keywords
TaqMan-based qPCR; Enterocytozoon hepatopenaei (EHP); Body mass index (BMI)
Categories
Funding
- China Agriculture Research System [CARS-47]
- Project of the Aoshan Sci. & Tech. Innovation Program of Qingdao National Laboratory for Marine Science and Technology [2015ASKJ02]
- Construction Programme for Taishan Scholars of Shandong Province
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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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