摘要
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.
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.
作者
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
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(Department of Molecular Biology and Biotechnology, Pan-African University Institute of Basic Sciences, Technology and Innovation, Nairobi, Kenya;Innovation, Technology Transfer Department, Kenya Medical Research Institute (ITTD-KEMRI), Nairobi, Kenya;Zoology Department, School of Biological Sciences, College of Pure and Applied Sciences, Jomo Kenyatta University of Agriculture and Technology, Nairobi, Kenya;Production Department, KEMRI, Nairobi, Kenya;Centre for Microbiology Research, Kenya Medical Research Institute, Nairobi, Kenya;Nagasaki University Institute of Tropical Medicine, Kenya Medical Research Institute (NUITM-KEMRI), Nairobi, Kenya;Department of Biochemistry, Microbiology and Biotechnology, Kenyatta University, Nairobi, Kenya;Department of Pharmacology and Pharmacognosy, University of Nairobi, Nairobi, Kenya)
