{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,21]],"date-time":"2026-08-21T12:27:43Z","timestamp":1787315263826,"version":"build-2736575974"},"reference-count":46,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2021,9,16]],"date-time":"2021-09-16T00:00:00Z","timestamp":1631750400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001411","name":"Indian Council of Medical Research","doi-asserted-by":"publisher","award":["6\/9-7 (155)\/2017-ECD-II"],"award-info":[{"award-number":["6\/9-7 (155)\/2017-ECD-II"]}],"id":[{"id":"10.13039\/501100001411","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Diagnostics"],"abstract":"<jats:p>To strengthen malaria surveillance, field-appropriate diagnostics requiring limited technical resources are of critical significance. Loop-mediated isothermal amplification (LAMP) based malaria diagnostic assays are potential point-of-care tests with high sensitivity and specificity and have been used in low-resource settings. Plasmodium vivax\u2013specific consensus repeat sequence (CRS)-based and Plasmodium falciparum\u2013specific 18S rRNA primers were designed, and a two-tube LAMP assay was developed. The diagnostic performance of a closed-tube LAMP assay and Loopamp\u2122 Malaria Detection (Pan\/Pf, Pv) kit was investigated using nested PCR confirmed mono- and co-infections of P. vivax and P. falciparum positive (n = 149) and negative (n = 67) samples. The closed-tube Pv LAMP assay showed positive amplification in 40 min (limit of detection, LOD 0.7 parasites\/\u00b5L) and Pf LAMP assay in 30 min (LOD 2 parasites\/\u00b5L). Pv LAMP and Pf LAMP demonstrated a sensitivity and specificity of 100% (95% CI, 95.96\u2013100% and 89.85\u2013100%, respectively). The LoopampTM Pan\/Pf Malaria Detection kit demonstrated a sensitivity and specificity of 100%, whereas LoopampTM Pv showed a sensitivity of 98.36% (95% CI, 91.28\u201399.71%) and specificity of 100% (95% CI, 87.54\u2013100%). The developed two-tube LAMP assay is highly sensitive (LOD \u2264 2 parasite\/\u00b5L), demonstrating comparable results with the commercial Loopamp\u2122 Malaria Detection (Pf\/pan) kit, and was superior in detecting the P. vivax co-infection that remained undetected by the Loopamp\u2122 Pv kit. The developed indigenous two-tube Pf\/Pv malaria detection can reliably be used for mass screening in resource-limited areas endemic for both P. falciparum and P. vivax malaria.<\/jats:p>","DOI":"10.3390\/diagnostics11091689","type":"journal-article","created":{"date-parts":[[2021,9,16]],"date-time":"2021-09-16T21:38:12Z","timestamp":1631828292000},"page":"1689","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Development of Two-Tube Loop-Mediated Isothermal Amplification Assay for Differential Diagnosis of Plasmodium falciparum and Plasmodium vivax and Its Comparison with Loopamp\u2122 Malaria"],"prefix":"10.3390","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0554-0017","authenticated-orcid":false,"given":"Mudsser","family":"Azam","sequence":"first","affiliation":[{"name":"Molecular Biology Laboratory, ICMR- National Institute of Pathology, New Delhi 110029, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kirti","family":"Upmanyu","sequence":"additional","affiliation":[{"name":"Molecular Biology Laboratory, ICMR- National Institute of Pathology, New Delhi 110029, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ratan","family":"Gupta","sequence":"additional","affiliation":[{"name":"Department of Paediatrics, VMMC and Safdarjung Hospital, New Delhi 110029, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Karugatharayil Sasi","family":"Sruthy","sequence":"additional","affiliation":[{"name":"Molecular Biology Laboratory, ICMR- National Institute of Pathology, New Delhi 110029, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Monika","family":"Matlani","sequence":"additional","affiliation":[{"name":"Department of Microbiology, VMMC and Safdarjung Hospital, New Delhi 110029, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Deepali","family":"Savargaonkar","sequence":"additional","affiliation":[{"name":"ICMR-National Institute of Malaria Research, New Delhi 110077, India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8094-4703","authenticated-orcid":false,"given":"Ruchi","family":"Singh","sequence":"additional","affiliation":[{"name":"Molecular Biology Laboratory, ICMR- National Institute of Pathology, New Delhi 110029, India"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,9,16]]},"reference":[{"key":"ref_1","unstructured":"Malaria (2020). 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