Evaluation of the sensitivity of a measles diagnostic real-time RT-PCR assay incorporating recently observed priming mismatch variants, 2024

Euro Surveill. 2024 Jul;29(28):2400410. doi: 10.2807/1560-7917.ES.2024.29.28.2400410.

Abstract

We investigated a variant of measles virus that encodes three mismatches to the reverse priming site for a widely used diagnostic real-time RT-PCR assay; reduction of sensitivity was hypothesised. We examined performance of the assay in context of the variant using in silico data, synthetic RNA templates and clinical specimens. Sensitivity was reduced observed at low copy numbers for templates encoding the variant sequence. We designed and tested an alternate priming strategy, rescuing the sensitivity of the assay.

Keywords: Measles; Measles Virus; N450; Paramyxovirus; Primer; RT-PCR; Real-Time PCR; Variant; rRT-PCR.

Publication types

  • Evaluation Study

MeSH terms

  • Humans
  • Measles virus* / genetics
  • Measles virus* / isolation & purification
  • Measles* / diagnosis
  • Measles* / virology
  • RNA, Viral* / genetics
  • Real-Time Polymerase Chain Reaction* / methods
  • Reverse Transcriptase Polymerase Chain Reaction* / methods
  • Sensitivity and Specificity*

Substances

  • RNA, Viral