Establishment of PCR Assay for Detecting of Lymantria dispar Multiple-embedded Nucleopolyhedrovirus (LdMNPV)

TAO Jing; HAN Chong-xuan; ZHANG Yong-an; WANG Yu-zhu; QU Liang-jian; WANG Qing-hua

A technical architecture used for the microdetermination of Lymantria dispar multiple-embedded nucleopolyhedrovirus was set up. Based on egt gene of LdMNPV, a pair of primers was designed with the sensitivity of 1fg·mL-1 DNA. By using this pair of primers, the partial sequences of egt gene were amplified from the eggs, larvae and pupae of the infected L. dispar, which verified the feasibility of this method. The partial sequences of egt gene were also amplified from the polyhedron suspension and the minimum amount of detection could be as low as 5 OBs·mL-1 of polyhedron. Morphology research indicated that the surface of a few of polyhedrons was rough pitted and released virions. It could be a reason why the target DNA fragment could be amplified from the virus suspension. So in the detection of LdMNPV, suspension after trituration, filtration and centrifugalism can be used as template.

1. College of Forestry, Northwest A & F University, Yangling 712100, Shaanxi, China

2. Research Institute of Forest Ecology, Environment and Protection, Chinese Academy of Forestry

Received Date: 2012-01-12

Available Online: 2012-10-11

Abstract: A technical architecture used for the microdetermination of Lymantria dispar multiple-embedded nucleopolyhedrovirus was set up. Based on egt gene of LdMNPV, a pair of primers was designed with the sensitivity of 1fg·mL-1 DNA. By using this pair of primers, the partial sequences of egt gene were amplified from the eggs, larvae and pupae of the infected L. dispar, which verified the feasibility of this method. The partial sequences of egt gene were also amplified from the polyhedron suspension and the minimum amount of detection could be as low as 5 OBs·mL-1 of polyhedron. Morphology research indicated that the surface of a few of polyhedrons was rough pitted and released virions. It could be a reason why the target DNA fragment could be amplified from the virus suspension. So in the detection of LdMNPV, suspension after trituration, filtration and centrifugalism can be used as template.

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Reference (17)

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Establishment of PCR Assay for Detecting of Lymantria dispar Multiple-embedded Nucleopolyhedrovirus (LdMNPV)

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