{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,26]],"date-time":"2025-10-26T21:19:01Z","timestamp":1761513541210,"version":"3.40.5"},"reference-count":32,"publisher":"American Society for Microbiology","issue":"1","license":[{"start":{"date-parts":[[2015,1,1]],"date-time":"2015-01-01T00:00:00Z","timestamp":1420070400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Antimicrob Agents Chemother"],"published-print":{"date-parts":[[2015,1]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            The resistance of\n            <jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">Plasmodium falciparum<\/jats:named-content>\n            to some antimalarial drugs is linked to single-nucleotide polymorphisms (SNPs). Currently, there are no methods for the identification of resistant parasites that are sufficiently simple, cheap, and fast enough to be performed at point-of-care, i.e., in local hospitals where drugs are prescribed. Primer extension methods (PEXT) were developed to identify 4 SNPs in\n            <jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">P. falciparum<\/jats:named-content>\n            positioned at amino acids 86, 184, and 1246 of the\n            <jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">P. falciparum<\/jats:named-content>\n            multidrug resistance 1 gene (\n            <jats:italic>pfmdr1<\/jats:italic>\n            ) and amino acid 76 of the chloroquine resistance transporter gene (\n            <jats:italic>pfcrt<\/jats:italic>\n            ). The PEXT products were visualized by a nucleic acid lateral flow immunoassay (NALFIA) with carbon nanoparticles as the detection labels. PCR-PEXT-NALFIAs showed good correlation to the reference methods, quantitative PCR (qPCR) or direct amplicon sequence analysis, in an initial open-label evaluation with 17 field samples. The tests were further evaluated in a blind study design in a set of 150 patient isolates. High specificities of 98 to 100% were found for all 4 PCR-PEXT genotyping assays. The sensitivities ranged from 75% to 100% when all PEXT-positive tests were considered. A number of samples with a low parasite density were successfully characterized by the reference methods but failed to generate a result in the PCR-PEXT-NALFIA, particularly those samples with microscopy-negative subpatent infections. This proof-of principle study validates the use of PCR-PEXT-NALFIA for the detection of resistance-associated mutations in\n            <jats:named-content xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" content-type=\"genus-species\" xlink:type=\"simple\">P. falciparum<\/jats:named-content>\n            , particularly for microscopy-positive infections. Although it requires a standard thermal cycler, the procedure is cheap and rapid and thus a potentially valuable tool for point-of-care detection in developing countries.\n          <\/jats:p>","DOI":"10.1128\/aac.03395-14","type":"journal-article","created":{"date-parts":[[2014,11,4]],"date-time":"2014-11-04T04:50:09Z","timestamp":1415076609000},"page":"365-371","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":13,"title":["Detection of Single-Nucleotide Polymorphisms in Plasmodium falciparum by PCR Primer Extension and Lateral Flow Immunoassay"],"prefix":"10.1128","volume":"59","author":[{"given":"A. P. H. A.","family":"Moers","sequence":"first","affiliation":[{"name":"Biomolecular Sensing &amp; Diagnostics, Food &amp; Biobased Research, Wageningen University and Research Centre, Wageningen, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"R. L.","family":"Hallett","sequence":"additional","affiliation":[{"name":"Faculty of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, London, United Kingdom"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"R.","family":"Burrow","sequence":"additional","affiliation":[{"name":"Faculty of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, London, United Kingdom"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"H. D. F. H.","family":"Schallig","sequence":"additional","affiliation":[{"name":"Koninklijk Instituut voor de Tropen\/Royal Tropical Institute, Biomedical Research, Amsterdam, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"C. J.","family":"Sutherland","sequence":"additional","affiliation":[{"name":"Faculty of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, London, United Kingdom"},{"name":"PHE Malaria Reference Laboratory, London School of Hygiene and Tropical Medicine, London, United Kingdom"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2502-5744","authenticated-orcid":false,"given":"A.","family":"van Amerongen","sequence":"additional","affiliation":[{"name":"Biomolecular Sensing &amp; Diagnostics, Food &amp; Biobased Research, Wageningen University and Research Centre, Wageningen, The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","volume-title":"Malaria fact sheet no. 94","author":"WHO","year":"2014","unstructured":"WHO. 2014. Malaria fact sheet no. 94. World Health Organization, Geneva, Switzerland. http:\/\/www.who.int\/mediacentre\/factsheets\/fs094\/en\/."},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.44.3.1087-1089.2006"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1186\/1475-2875-7-70"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1179\/136485907X156997"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1038\/35002615"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1093\/infdis\/jiu358"},{"key":"e_1_3_2_8_2","doi-asserted-by":"publisher","DOI":"10.1186\/1475-2875-3-9"},{"key":"e_1_3_2_9_2","doi-asserted-by":"publisher","DOI":"10.4269\/ajtmh.2002.67.578"},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.00875-06"},{"key":"e_1_3_2_11_2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.genom.2.1.235"},{"key":"e_1_3_2_12_2","doi-asserted-by":"publisher","DOI":"10.1038\/35103535"},{"key":"e_1_3_2_13_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00216-007-1593-4"},{"key":"e_1_3_2_14_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00216-007-1383-z"},{"key":"e_1_3_2_15_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.bios.2009.03.010"},{"key":"e_1_3_2_16_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00216-011-4845-2"},{"key":"e_1_3_2_17_2","doi-asserted-by":"crossref","first-page":"164","DOI":"10.1093\/clinchem\/47.2.164","article-title":"Enabling large-scale pharmacogenetic studies by high-throughput mutation detection and genotyping technologies","volume":"47","author":"Shi MM","year":"2001","unstructured":"Shi MM. 2001. Enabling large-scale pharmacogenetic studies by high-throughput mutation detection and genotyping technologies. Clin Chem 47:164\u2013172.","journal-title":"Clin Chem"},{"key":"e_1_3_2_18_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00216-010-4334-z"},{"key":"e_1_3_2_19_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00216-011-5340-5"},{"key":"e_1_3_2_20_2","doi-asserted-by":"crossref","first-page":"105","DOI":"10.3920\/978-90-8686-538-3","volume-title":"Rapid methods for biological and chemical contaminants in food and feed","author":"van Amerongen A","year":"2005","unstructured":"van Amerongen A, Koets M. 2005. Simple and rapid bacterial protein and DNA diagnostic methods based on signal generation with colloidal carbon particles, p 105\u2013126. In van Amerongen A, Barug D, Lauwaars (ed), Rapid methods for biological and chemical contaminants in food and feed. Wageningen Academic Publishers, Wageningen, The Netherlands."},{"key":"e_1_3_2_21_2","doi-asserted-by":"publisher","DOI":"10.1373\/clinchem.2007.102491"},{"key":"e_1_3_2_22_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jim.2011.11.003"},{"key":"e_1_3_2_23_2","doi-asserted-by":"publisher","DOI":"10.1186\/1475-2875-11-279"},{"key":"e_1_3_2_24_2","doi-asserted-by":"publisher","DOI":"10.1093\/jac\/dkm104"},{"key":"e_1_3_2_25_2","doi-asserted-by":"publisher","DOI":"10.1186\/1475-2875-9-74"},{"key":"e_1_3_2_26_2","doi-asserted-by":"publisher","DOI":"10.2144\/97233rr01"},{"key":"e_1_3_2_27_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.aca.2007.07.011"},{"key":"e_1_3_2_28_2","doi-asserted-by":"publisher","DOI":"10.1021\/ac201823v"},{"key":"e_1_3_2_29_2","doi-asserted-by":"publisher","DOI":"10.1007\/s00216-013-7192-7"},{"key":"e_1_3_2_30_2","first-page":"185","article-title":"NASBA and other transcription-based amplification methods for research and diagnostic microbiology","volume":"10","author":"Chan AB","year":"1999","unstructured":"Chan AB, Fox JD. 1999. NASBA and other transcription-based amplification methods for research and diagnostic microbiology. Rev Med Microbiol 10:185\u2013196.","journal-title":"Rev Med Microbiol"},{"key":"e_1_3_2_31_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/28.12.e63"},{"key":"e_1_3_2_32_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/20.7.1691"},{"key":"e_1_3_2_33_2","doi-asserted-by":"publisher","DOI":"10.1586\/14737159.2.6.542"}],"container-title":["Antimicrobial Agents and Chemotherapy"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/AAC.03395-14","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/AAC.03395-14","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,2,21]],"date-time":"2022-02-21T22:53:36Z","timestamp":1645484016000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/AAC.03395-14"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2015,1]]},"references-count":32,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2015,1]]}},"alternative-id":["10.1128\/AAC.03395-14"],"URL":"https:\/\/doi.org\/10.1128\/aac.03395-14","relation":{},"ISSN":["0066-4804","1098-6596"],"issn-type":[{"type":"print","value":"0066-4804"},{"type":"electronic","value":"1098-6596"}],"subject":[],"published":{"date-parts":[[2015,1]]},"assertion":[{"value":"2014-05-20","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2014-10-23","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2014-12-23","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}