A proposed validation method for automated nucleic acid extraction and RT-qPCR analysis: an example using Bluetongue virus.

Last updated on 2-10-2019 by Anonymous (not verified)

Peer reviewed scientific article

SCIENSANO

Abstract:

This study proposes a validation strategy for an automated extraction procedure, followed by RT-qPCR analysis. To avoid false-negative results, a triplex RT-qPCR was used which detects the target viral RNA, an internal and an external control. The methods to determine the validation parameters such as linearity, efficiency, analytical sensitivity, analytical specificity and intra- and interrun variability are described in detail. Special attention is given to the analytical sensitivity, which is determined by probit analysis. The limit of detection was set at the input concentration resulti…

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