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Analysis of drug resistance marker genes of Plasmodium falciparum after implementation of artemisinin-based combination therapy in Pune district, India

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Abstract

The global emergence and spread of malaria parasites resistant to antimalarial drugs is a major problem in malaria control and elimination. In this study, samples from Pune district were characterized to determine prevalence of molecular markers of resistance to chloroquine (pfcrt codons C72S, M74I, N75E, K76T and pfmdr-1 N86Y, Y184F), pyrimethamine (pfdhfr C50R, N51I, C59R, S108N), sulfadoxine (pfdhps, S436A, A437G, K540E, A581G), and artemisinin (pfkelch13, C580Y, R539T). The pfcrt K76T mutation was found in 78% samples as CVMNT, SVMNT and CVIET haplotype. The pfmdr-1 N86Y and Y184F mutations were found in 54% of samples. The pfdhfr double mutation C59R + S108N was present in 67% of samples, while the pfdhfr triple mutation (N51I + C59R + S108N) was not detected. The pfdhps mutations A437G and K540E were found in 67% of samples. Single mutants of pfdhps were rare, with K540E detected in only 6 patient samples. Similarly, pfdhps A581G was found in 13 of the isolates. The molecular markers associated with artemisinin resistance (mutations in pfkelch13 C580Y, R539T) were not detected in any of the isolates. These results suggest an emerging problem with multidrug-resistant P. falciparum. Though the genotype conventionally associated with artemisinin resistance was not observed, chloroquine-resistant genotype has reached complete fixation in the population. Moreover, the prevalence of mutations in both pfdhfr and pfdhps, with the presence of the quadruple mutant, indicates that continued monitoring is required to assess whether sulfadoxine-pyrimethamine can be used efficiently as a partner drug for artemisinin for the treatment of P. falciparum.

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Abbreviations

ACT:

Artemisinin-based combination therapy

BLAST:

Basic Local Alignment Search Tool

DNA:

Deoxyribonucleic acid

MSP:

Merozoite surface protein

PCR:

Polymerase chain reaction

pfcrt :

Plasmodium falciparum Chloroquine resistance transporter

pfdhfr :

Plasmodium falciparum Dihydrofolate reductase

pfdhps :

Plasmodium falciparum Dihydropteroate synthase

pfmdr-1 :

Plasmodium falciparum Multidrug resistance protein 1

RSA:

Ring-stage survival assay

SNP:

Single nucleotide polymorphism

SP:

Sulfadoxine-pyrimethamine

WHO:

World Health Organisation

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Acknowledgements

This work is supported by the DST WOS A Grant (SR/WOS-A/LS-146/2011) to AO. KK is supported by the Genome Engineering Technologies program (BT/PR25858/GET/119/169/2017) of the Department of Biotechnology (DBT) from the Government of India. The funders had no role in study design, data collection, and analysis, decision to publish, or preparation of the manuscript. We would like to thank Dr Tushar Patil, Dr Ashwin Khandare and Mr Mangesh Deval for helping AO with sample collection. We thank Sarthak Joshi for the structural modelling and SNP mapping.

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Authors and Affiliations

  1. Department of Zoology, Centre for Advanced Studies, Savitribai Phule Pune University, Pune, India

    Aarti Ozarkar, Swati Dass & Deepti Deobagkar

  2. National Centre for Cell Science, Pune, India

    Prakash Deshpande

  3. Division of Biology, Indian Institute of Science Education and Research (IISER), Pune, India

    Abhishek Kanyal & Krishanpal Karmodiya

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  1. Aarti Ozarkar
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  2. Abhishek Kanyal
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  3. Swati Dass
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Corresponding authors

Correspondence to Aarti Ozarkar or Krishanpal Karmodiya.

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The ethics Committee and the Institutional Review Board of Pune University, Pune, Maharashtra, India, has approved this study. Informed written consent was obtained from all the study participants.

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Corresponding editor: BJ RAO

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Ozarkar, A., Kanyal, A., Dass, S. et al. Analysis of drug resistance marker genes of Plasmodium falciparum after implementation of artemisinin-based combination therapy in Pune district, India. J Biosci 46, 77 (2021). https://doi.org/10.1007/s12038-021-00200-3

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  • DOI: https://doi.org/10.1007/s12038-021-00200-3

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