Development of a Real-Time RT-PCR Method for Detection of Chronic Bee Paralysis Virus
To establish a rapid diagnostic method for chronic bee paralysis virus(CBPV),a fluorogenic TaqMan real-time PCR assay was developed by designing specific primers and TaqMan probes according to the RNA dependent RNA polymerase(RdRp)gene sequence of CBPV in this study.The results showed that the real-time RT-PCR method had a good linear relation,and linear correlation coefficient R2 was 0.998.The sensitivity limit was 10 copies/μL and there was no cross-reaction with other Honey bee virus,which indicated that the method had good sensitivity and specificity.The coefficients of variation(CV)for intra-assay and inter-assay repeatability were less than 0.5％and 2％,respectively,showing that the method had good stability.The real-time RT-PCR method for CBPV developed in this study might be used for laboratory testing,epidemiological investigation and epidemic monitoring.
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