A LAMP-SNP Assay Detecting C580Y Mutation in Pfkelch13 Gene from Clinically Dried Blood Spot Samples

Thunchanok Khammanee; Nongyao Sawangjaroen; Hansuk Buncherd; Aung Win Tun; Supinya Thanapongpichat

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자료유형: 학술저널

저자정보: Thunchanok Khammanee (Prince of Songkla University) Nongyao Sawangjaroen (Prince of Songkla University) Hansuk Buncherd (Prince of Songkla University) Aung Win Tun (Mahidol University) Supinya Thanapongpichat (Prince of Songkla University)

저널정보: 대한기생충학열대의학회 Parasites, Hosts and Diseases The Korean Journal of Parasitology Vol.59 No.1

발행연도: 2021.2

수록면: 15 - 22 (8page)

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Artemisinin resistance (ART) has been confirmed in Greater Mekong Sub-region countries. Currently, C580Y mutation on Pfkelch13 gene is known as the molecular marker for the detection of ART. Rapid and accurate detection of ART in field study is essential to guide malaria containment and elimination interventions. A simple method for collection of malaria-infected blood is to spot the blood on filter paper and is fast and easy for transportation and storage in the field study. This study aims to evaluate LAMP-SNP assay for C580Y mutation detection by introducing an extra mismatched nucleotide at the 3’ end of the FIP primer. The LAMP-SNP assay was performed in a water bath held at a temperature of 56°C for 45 min. LAMP-SNP products were interpreted by both gel-electrophoresis and HNB-visualized changes in color. The method was then tested with 120 P. falciparum DNA from dried blood spot samples. In comparing the LAMP-SNP assay results with those from DNA sequencing of the clinical samples, the 2 results fully agreed to detect C580Y. The sensitivity and specificity of the LAMP-SNP assay showed 100%. There were no cross-reactions with other Plasmodium species and other Pfkelch13 mutations. The LAMP-SNP assay performed in this study was rapid, reliable, and useful in detecting artemisinin resistance in the field study.

#Plasmodium falciparum #Pfkelch13 gene #artemisinin resistance #C580Y mutation #LAMP-SNP assay

참고문헌 (19)

참고문헌 신청

World Malaria Report 2018

Dondorp AM / 200 / Artemisinin resistance in Plasmodium falciparum malaria / N Engl J Med 361:455~467

Ariey F / 2014 / A molecular marker of artemisinin-resistant Plasmodium falciparum malaria / Nature 505:50~55

Witkowski B / 2013 / Novel phenotypic assays for the detection of artemisinin-resistant Plasmodium falciparum malaria in Cambodia: in-vitro and ex-vivo drug-response studies / Lancet Infect Dis 13:1043~1049

Judith Straimer / 2015 / K13-propeller mutations confer artemisinin resistance in Plasmodium falciparum clinical isolates / Science 347(6220):428~431

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