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

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

Andrew S Beck  1 , Elena N Lopareva  1 , Hyun Hwang  2 , Derek Hart  3 , Marcos de Almeida  4 , Raydel Anderson  1 , Paul A Rota  1 , Bettina Bankamp  1
Affiliations

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

Andrew S Beck et al. Euro Surveill. 2024 Jul.

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.

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Conflict of interest statement

Conflict of interest: None declared.

Figures

Figure 1
Figure 1
Measles virus RT-rPCR assay primer/probe sequences and 3UC variant surveillance data, 2003–2023 (n = 37,995)
Figure 2
Figure 2
Summary of measles virus RT-rPCR sensitivities using synthetic RNA templates and clinical specimens
See this image and copyright information in PMC

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