Research on the detection of bee viruses by one step and two step RT-PCR
Author of the article:JIA Hui-Ru1** ZHOU Ting1 GAO Fu-Chao2***
Author's Workplace：1. Key Laboratory of Pollinating Insect Biology of Agriculture, Institute of Apiculture Research, Chinese Academy of Agriculture Science, Beijing 100093, China; 2. Heilongjiang Academy of Agricultural Sciences, Mudanjiang, 157041, China
Key Words： bee virus, one-step RT-PCR, two-step RT-PCR
Abstract：
[Objectives] To find which RT-PCR technology can effectively detect bee viruses. [Methods] Bee viruses (IAPV, DWV, SBV, ABPV, BQCV, and CBPV) were extracted from bees and the sensitivity of one and two step RT-PCR in detecting these six virus was compared. [Results] Both methods amplified viral fragments, which were 158 (IAPV), 269 (DWV), 342 (SBV), 460 (ABPV), 536 (BQCV) and 774 bp (CBPV), in length. Serial dilutions assay showed that two-step RT-PCR detected much lower concentrations of virus particles. [Conclusion] Comparison of one-step and two-step RT-PCR indicates that both methods can effectively detect many bee viruses under optimized conditions, however, two-step RT-PCR is more sensitive than one-step RT-PCR.