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Sensitive poliovirus detection using nested PCR and nanopore sequencing: a prospective validation study
Article de revue scientifique   Open Access   Avec comité de lecture

Sensitive poliovirus detection using nested PCR and nanopore sequencing: a prospective validation study

Alexander Shaw, Tresor Kabeya Mampuela, Emmanuel Lokilo Lofiko, Catherine Pratt, Catherine Troman, Erika Bujaki, Áine O’toole, Joyce Odeke Akello, Adrienne Amuri Aziza, Eddy Kinganda Lusamaki, …
Nature Microbiology, Vol.8(9), p.1634-1640
17/08/2023

Résumé

Dansyl Compounds Nanopore Sequencing* Poliovirus* / genetics Polymerase Chain Reaction
Timely detection of outbreaks is needed for poliovirus eradication, but gold standard detection in the Democratic Republic of the Congo takes 30 days (median). Direct molecular detection and nanopore sequencing (DDNS) of poliovirus in stool samples is a promising fast method. Here we report prospective testing of stool samples from suspected polio cases, and their contacts, in the Democratic Republic of the Congo between 10 August 2021 and 4 February 2022. DDNS detected polioviruses in 62/2,339 (2.7%) of samples, while gold standard combination of cell culture, quantitative PCR and Sanger sequencing detected polioviruses in 51/2,339 (2.2%) of the same samples. DDNS provided case confirmation in 7 days (median) in routine surveillance conditions. DDNS enabled confirmation of three serotype 2 circulating vaccine-derived poliovirus outbreaks 23 days (mean) earlier (range 6–30 days) than the gold standard method. The mean sequence similarity between sequences obtained by the two methods was 99.98%. Our data confirm the feasibility of implementing DDNS in a national poliovirus laboratory.

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