A novel real-time PCR approach for detection of infectious hypodermal and haematopoietic necrosis virus (IHHNV) in the freshwater crayfish Procambarus clarkii

作者：Chen, Bo-Kun; Dong, Zhen; Pang, Neng-Yuan; Nian, Yan-Ying; Yan, Dong-Chun^*

来源：Journal of Invertebrate Pathology, 2018, 157: 100-103.

DOI：10.1016/j.jip.2018.08.009

• Real-time PCR
• Infectious hypodermal and haematopoietic necrosis virus (IHHNV)
• Procambarus clarkii

Infectious hypodermal and haematopoietic necrosis virus (IHHNV) infects many crustacean hosts, including cultured penaeid shrimp. In the present study, we aimed to develop a novel sensitive SYBR Green-based real-time PCR method to specifically amplify DNA fragments of IHHNV. Our newly developed real-time PCR method with a 195-bp amplicon specifically detected IHHNV and showed no cross reaction with white spot syndrome virus (WSSV), hepatopancreatic parvovirus (HPV), Enterocytozoon hepatopenaei (EHP), infectious myonecrosis virus (IMNV) and yellow-head virus (YHV). This method could detect as low as one single copy of IHHNV plasmid DNA, more sensitive than other SYBR Green-based real-time PCR methods and less expensive and more convenient than the TaqMan probe-based real-time PCR. Moreover, our data using the newly designed method showed that 80% of IHHNV-fed Procambarus clarkii samples were IHHNV positive. Our findings further confirmed that P. clarkii can be infected by IHHNV.

• 出版日期2018-9
• 单位山东农业大学; 鲁东大学