Short communication
Capillary electrophoresis of PCR fragments with 5´-labelled primers for testing the SARS-Cov-2

https://doi.org/10.1016/j.jviromet.2020.113937Get rights and content

Highlights

  • We designated an alternative to qtPCR for the identification of SARS-Cov-2.

  • The method was a PCR with 5-end fluorescent primers and capillary electrophoresis.

  • Two viral fragments amplified in several SARS-Cov-2 positive and negative specimens.

  • This approach would increase the testing capacity of diagnostic labs.

Abstract

Due to the huge demand for SARS-Cov-2 determination,alternatives to the standard qtPCRtestsare potentially useful for increasing the number of samples screened. Our aim was to develop a direct fluorescent PCR capillary-electrophoresis detection of the viral genome. We validated this approach on several SARS-Cov-2 positive and negative samples.We isolated the naso-pharingealRNA from 20 positive and 10 negative samples. The cDNA was synthesised and two fragments of the SARS-Cov-2 were amplified. One of the primers for each pair was 5´-end fluorochrome labelled. The amplifications were subjected to capillary electrophoresis in ABI3130 sequencers to visualize the fluorescent peaks.The two SARS-Cov-2 fragments were successfully amplified in the positive samples, while the negative samples did not render fluorescent peaks. In conclusion, we describe and alternative method to identify the SARS-Cov-2 genome that could be scaled to the analysis of approximately 100 samples in less than 5 h. By combining a standard PCR with capillary electrophoresis our approach would overcome the limits imposed to many labs by the qtPCR and increase the testing capacity.

Keywords

COVID19
SARS-Cov-2
Viral genome detection
Quantitative PCR
Capillary electrophoresis

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