Investigation of Plasmodium falciparum Resistance Biomarkers among Primary School Children in Western Kenya

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

作者机构：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 

出 版 物：《American Journal of Molecular Biology》 (美国分子生物学期刊（英文）)

年 卷 期：2022年第12卷第2期

页      面：43-53页

学科分类：1002[医学-临床医学] 100201[医学-内科学(含：心血管病、血液病、呼吸系病、消化系病、内分泌与代谢病、肾病、风湿病、传染病)] 10[医学] 

主　　题：P. falciparum Chloroquine Drug Resistance Malaria P. falciparum Chloroquine Resistance Transporter Gene Mutations 

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 *** to Pfcrt which calls for robust continuous monitoring and surveillance. Conclusion: Due to the increase of the resistant Pfcrt gene pre