PropertyValue
?:abstract
  • Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged in December 2019 and quickly spread around the world, forcing global health authorities to develop protocols for its diagnosis. Here we report dimer formation in the N2 primers-probe set (CDC 2019-nCoV Real-Time RT-PCR) used in the diagnostic routine, and propose alternatives to reduce dimerization events. Late unspecific amplifications were visualized in 56.4% of negative samples and 57.1% of no-template control, but not in positive samples or positive control. In silico analysis and gel electrophoresis confirmed the dimer formation. The RT-qPCR parameters were optimized and the late unspecific amplifications decreased to 11.5% in negative samples and no-template control. The adjustment of PCR parameters was essential to reduce the risk of false-positives results and to avoid inclusive results requiring repeat testing, which increases the costs and generates delays in results or even unnecessary requests for new samples.
is ?:annotates of
?:creator
?:journal
  • Int_J_Infect_Dis
?:license
  • unk
?:publication_isRelatedTo_Disease
?:source
  • WHO
?:title
  • Adjusting RT-qPCR conditions to avoid unspecific amplification in SARS-CoV-2 diagnosis
?:type
?:who_covidence_id
  • #898983
  • #997013
?:year
  • 2020

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