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Development and Evaluation of a Novel HNB Based Isothermal Amplification Assay for Fast Detection of Pyrimethamine Resistance (S108N) in Plasmodium falciparum

Author

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  • Madhvi Chahar

    (Division of Epidemiology and Clinical Research, National Institute of Malaria Research, Sector-8 Dwarka, New Delhi 110077, India)

  • Anup Anvikar

    (Division of Epidemiology and Clinical Research, National Institute of Malaria Research, Sector-8 Dwarka, New Delhi 110077, India)

  • Neena Valecha

    (Division of Epidemiology and Clinical Research, National Institute of Malaria Research, Sector-8 Dwarka, New Delhi 110077, India)

Abstract

Sulphadoxine and pyrimethamine (SP) have been used as long-acting partner antimalarial drugs in artemisinin combination therapy (ACT) for falciparum malaria. The emergence and increasing spread of SP resistance in malaria-endemic areas have become a challenge for the control of malaria. Therefore, regular monitoring of the mutation status of partner drugs is important for the better management of drug policy. There are limitations with traditional molecular methods and there is an urgent need for an easy method for diagnosis of drug resistance. In this study we have introduced and developed a novel single nucleotide polymorphism loop-mediated isothermal amplification (SNP–LAMP) approach based on a hydroxynaphthol blue (HNB) indicator for the easier and quicker detection of pyrimethamine resistance in Plasmodium falciparum malaria. To implement this novel approach, many sets of LAMP primers were designed and tested. Finally, one set of forward inner primer M1 (FIPM1) of LAMP primer was selected that specifically distinguishes pyrimethamine-resistant P. falciparum malaria. The LAMP reactions were optimized at 60–66 °C for 45 min. High sensitivity (7 parasites/µL) was observed with 10 −4 fold dilutions (<2 ng DNA) of genomic DNA. Moreover, this approach has the potential to be applied even in laboratories unfamiliar with PCR or other molecular methods, and in future, this can be helpful for the better management of anti-malarial policy.

Suggested Citation

  • Madhvi Chahar & Anup Anvikar & Neena Valecha, 2019. "Development and Evaluation of a Novel HNB Based Isothermal Amplification Assay for Fast Detection of Pyrimethamine Resistance (S108N) in Plasmodium falciparum," IJERPH, MDPI, vol. 16(9), pages 1-13, May.
  • Handle: RePEc:gam:jijerp:v:16:y:2019:i:9:p:1635-:d:229935
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    References listed on IDEAS

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    1. Frédéric Ariey & Benoit Witkowski & Chanaki Amaratunga & Johann Beghain & Anne-Claire Langlois & Nimol Khim & Saorin Kim & Valentine Duru & Christiane Bouchier & Laurence Ma & Pharath Lim & Rithea Lea, 2014. "A molecular marker of artemisinin-resistant Plasmodium falciparum malaria," Nature, Nature, vol. 505(7481), pages 50-55, January.
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