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Deletions of the\n                    <jats:italic>hrp2<\/jats:italic>\n                    and\n                    <jats:italic>hrp3<\/jats:italic>\n                    genes result in false-negative tests and are a threat for malaria control. A novel assay for molecular surveillance of\n                    <jats:italic>hrp2<\/jats:italic>\n                    \/\n                    <jats:italic>hrp3<\/jats:italic>\n                    deletions was developed based on droplet digital PCR (ddPCR). The assay quantifies\n                    <jats:italic>hrp2<\/jats:italic>\n                    ,\n                    <jats:italic>hrp3<\/jats:italic>\n                    , and a control gene with very high accuracy. The theoretical limit of detection was 0.33 parasites\/\u00b5l. The deletion was reliably detected in mixed infections with wild-type and\n                    <jats:italic>hrp2<\/jats:italic>\n                    -deleted parasites at a density of &gt;100 parasites\/reaction. For a side-by-side comparison with the conventional nested PCR (nPCR) assay, 248 samples were screened in triplicate by ddPCR and nPCR. No deletions were observed by ddPCR, while by nPCR\n                    <jats:italic>hrp2<\/jats:italic>\n                    deletion was observed in 8% of samples. The ddPCR assay was applied to screen 830 samples from Kenya, Zanzibar\/Tanzania, Ghana, Ethiopia, Brazil, and Ecuador. Pronounced differences in the prevalence of deletions were observed among sites, with more\n                    <jats:italic>hrp3<\/jats:italic>\n                    than\n                    <jats:italic>hrp2<\/jats:italic>\n                    deletions. In conclusion, the novel ddPCR assay minimizes the risk of false-negative results (i.e.,\n                    <jats:italic>hrp2<\/jats:italic>\n                    deletion observed when the sample is wild type), increases sensitivity, and greatly reduces the number of reactions that need to be run.\n                  <\/jats:p>","DOI":"10.7554\/elife.72083","type":"journal-article","created":{"date-parts":[[2022,6,7]],"date-time":"2022-06-07T07:15:10Z","timestamp":1654586110000},"update-policy":"https:\/\/doi.org\/10.7554\/elife.72083","source":"Crossref","is-referenced-by-count":58,"title":["High-throughput Plasmodium falciparum hrp2 and hrp3 gene deletion typing by digital PCR to monitor malaria rapid diagnostic test efficacy"],"prefix":"10.7554","volume":"11","author":[{"given":"Claudia A","family":"Vera-Arias","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00mkhxb43","id-type":"ROR","asserted-by":"publisher"}],"name":"University of Notre Dame","place":["Notre 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