TITLE:
Investigation of Plasmodium falciparum Resistance Biomarkers among Primary School Children in Western Kenya
AUTHORS:
Otilmoi Poul Stephen, Tonny Teya Nyandwaro, Robinson Mugasiali Irekwa, Rebecca Wanjiku Waihenya, Matthew Mutinda Munyao, Peter Kipkemboi Rotich, Caroline Wangui Njoroge, Anne Wanjiru Mwangi, Joanne Jepkemei Yego, Nicole Sian Tanchu, Dawala Koromtili Oumar, Grace Ngendo Kanyita, Primrose Muthoni Ndungu, Samson Muuo Nzou
KEYWORDS:
P. falciparum, Chloroquine, Drug Resistance, Malaria, P. falciparum Chloroquine Resistance Transporter Gene, Mutations
JOURNAL NAME:
American Journal of Molecular Biology,
Vol.12 No.2,
April
26,
2022
ABSTRACT: Background: A marked decrease in malaria-related deaths worldwide has been
attributed to the administration of effective antimalarials against Plasmodium
falciparum. However, the continuous spread of P. falciparum resistance to anti-malarial drugs is raising a serious problem in controlling
Malaria to the vulnerable children’s immune system. In recent studies, Plasmodium falciparum Kelch 13 propeller
gene (Pfk13) has been reported to
develop resistance to artemisinin in South Asia. In this study, we checked Plasmodium falciparum chloroquine
resistance transporter gene (Pfcrt)
involved in chloroquine (CQ) resistance. Method: In this study, archived
280 samples were collected from Alupe primary school children in Busia, Western
Kenya from May, 2016 to November, 2016. Genomic DNA was extracted using the
MightyPrep reagent. The samples were investigated for P. falciparum positivity out of which 67 of them tested positive
giving a prevalence rate of 24%. The sixty-seven were subjected to PCR
amplification for the molecular marker resistance to Pfcrt. After PCR
amplification, the amplicons were purified and sequenced using Sanger
Sequencing. The sequence data were analyzed using BioEdit software to identify
point mutations. Results: 14 samples sequences were analyzed on Bioedit
software giving the following amino acid changes F76C, Y66H, L70A, Y58C, T59V,
V65I, P67L, T81L, Y60S, Y66S, P67T and I71F).
New mutations have been reported at position 76 leading to an amino acid
change, one of Pfcrt gold standard biomarkers. However,
amino acid changes Y66H, L70A, Y58C, T59V, V65I, P67L, T81L, Y60S, Y66S, P67T
and I71F are newly reported giving an increase in Pfcrt prevalence of concern
from zero to 5.0%. A phylogenetic evolutionary relationship was constructed as
shown below. Generally, the results showed a continuous resistance of P.falciparum to Pfcrt which calls for robust continuous
monitoring and surveillance. Conclusion: Due to the increase of the
resistant Pfcrt gene prevalence, continuous development of new mutants against
chloroquine indicates that there is need to repurpose anti-malarial drugs for
future partner drugs.