Validation of quantitative real-time RT-PCR assays for the detection of six honeybee viruses.

Validation of quantitative real-time RT-PCR assays for the detection of six honeybee viruses.
J Virol Methods. 2019 Apr 23;:
Authors: Schurr F, Tison A, Militano L, Cheviron N, Sircoulomb F, Rivière MP, Ribière-Chabert M, Thiéry R, Dubois E
Abstract
Acute bee paralysis virus (ABPV), Black queen cell virus (BQCV), Chronic bee paralysis virus (CBPV), Deformed wing virus (DWV), Sacbrood virus (SBV) and Varroa destructor virus-1 (VDV1) are the six main honeybee viruses reported in Europe. We assessed the accuracy (trueness and precision) of reverse transcriptase quantitative TaqMan® PCR methods (RT-qPCR) for quantifying ABPV, BQCV, DWV, VDV1 and SBV loads. Once the systematic bias in quantitative results had been corrected (overestimation in ABPV and BQCV quantification and underestimation in that of SBV and VDV1), measurements were taken to determine the viral load ranges for which quantification uncertainty was below ± 1 log10 equivalent of genome copies per bee (hereafter reported as genome copies/bee). The accuracy range of RT-qPCR was found to be between 6.4 and 10.4 log10 genome copies/bee for ABPV, between 3.0 and 10.0 log10 genome copies/bee for BQCV, between 2.4 and 10.4 log10 genome copies/bee for DWV and between 3.4 and 10.4 log10 genome copies/bee for SBV. Outside these ranges, the results' uncertainty is higher. VDV1 RT-qPCR accuracy was outside validation limits for all viral loads. Using these RT-qPCR methods, we quan...

Source: Journal of Virological Methods - April 22, 2019 Category: Virology Authors: Schurr F, Tison A, Militano L, Cheviron N, Sircoulomb F, Rivière MP, Ribière-Chabert M, Thiéry R, Dubois E Tags: J Virol Methods Source Type: research