Jian Li1, Jiangtao Chen2, Dongde Xie2, Urbano Monsuy Eyi3, Rocio Apicante Matesa3, Maximo Miko Ondo Obono3, Carlos Sala Ehapo3, Liye Yang4, Huitian Yang4, Min Lin5, Wanjun Wu1, Kai Wu6, Shan Li1, Zongyun Chen1. 1. Department of Parasitology, College of Basic Medicine, Department of Infectious Diseases, Shiyan Renmin Hospital, Hubei University of Medicine, Shiyan 442000, People's Republic of China. 2. The Chinese Medical Aid Team to the Republic of Equatorial Guinea, Guangzhou 510000, People's Republic of China. 3. Central Blood Transfusion Service, Department of Medical Laboratory Science, Malabo Regional Hospital, Malabo, Equatorial Guinea. 4. Laboratory Medical Center, Chaozhou Central Hospital, Southern Medical University, Chaozhou 521021, People's Republic of China. 5. Laboratory Medical Center, Chaozhou Central Hospital, Southern Medical University, Chaozhou 521021, People's Republic of China. Electronic address: Konfutea@hotmail.com. 6. Department of Schistosomiasis and Endemic Diseases, Wuhan City Center for Disease Control and Prevention, Wuhan 430015, People's Republic of China.
Abstract
BACKGROUND: Antimalarial drug resistance is a primary public health problem. Haplotypes of pfcrt and pfmdr1 gene have been implicated to be molecular markers of chloroquine (CQ) resistance. This study aims to explore mutation distribution of Pfcrt and Pfmdr1 in Bioko Island, Equatorial Guinea (EG). METHODS: Blood samples were collected from different districts of Bioko. The single nucleotide polymorphisms in Pfcrt (codons 72 to 76) and Pfmdr1 (codons 86, 130, 184, 1034, 1042, 1109 and 1246) were assessed by nested PCR with DNA sequencing and haplotype prevalences were also determined. RESULTS: Analysis of Pfcrt and Pfmdr1 mutations was successful in 151 and 157 samples respectively out of the 172 samples taken for this study. The mutations of Pfcrt and Pfmdr1 were found in 98.67% and 89.81% isolates, respectively. The Pfcrt 74-76, Pfmdr1 86, and Pfmdr1 184 were 92.05%, 50.32%, and 87.26% found mostly of mutation type, respectively. Three haplotypes coding 72-76 of Pfcrt were found including CVMNK, CVIET, and CVM/I N/E K/T, which accounted for 1.33%, 92.05%, and 6.62%, respectively. No mutation in Pfmdr1-N1 codon at 130 and Pfmdr1-N2 (S1034C, N1042D, V1109I, and D1246Y) was detected. The types coding 86 and 184 in Pfmdr1 were found including NY, YY, NF, YF, NY/F and YY/F, which accounted for 10.19%, 2.55%, 33.76%, 45.22%, 5.73% and 2.55%, respectively. CONCLUSION: High prevalence of Pfcrt CVIET and Pfmdr1 86Y, 184F double mutations confirm high-level CQ resistance (CQR) and might suggest reduced susceptibility of Plasmodium falciparum isolates to AQ in Bioko, EG. It establishes fundamental data for detection of P. falciparum CQR with molecular markers and will promotes the surveillance level of drug resistance in Bioko, EG.
BACKGROUND: Antimalarial drug resistance is a primary public health problem. Haplotypes of pfcrt and pfmdr1 gene have been implicated to be molecular markers of chloroquine (CQ) resistance. This study aims to explore mutation distribution of Pfcrt and Pfmdr1 in Bioko Island, Equatorial Guinea (EG). METHODS: Blood samples were collected from different districts of Bioko. The single nucleotide polymorphisms in Pfcrt (codons 72 to 76) and Pfmdr1 (codons 86, 130, 184, 1034, 1042, 1109 and 1246) were assessed by nested PCR with DNA sequencing and haplotype prevalences were also determined. RESULTS: Analysis of Pfcrt and Pfmdr1 mutations was successful in 151 and 157 samples respectively out of the 172 samples taken for this study. The mutations of Pfcrt and Pfmdr1 were found in 98.67% and 89.81% isolates, respectively. The Pfcrt 74-76, Pfmdr1 86, and Pfmdr1 184 were 92.05%, 50.32%, and 87.26% found mostly of mutation type, respectively. Three haplotypes coding 72-76 of Pfcrt were found including CVMNK, CVIET, and CVM/I N/E K/T, which accounted for 1.33%, 92.05%, and 6.62%, respectively. No mutation in Pfmdr1-N1 codon at 130 and Pfmdr1-N2 (S1034C, N1042D, V1109I, and D1246Y) was detected. The types coding 86 and 184 in Pfmdr1 were found including NY, YY, NF, YF, NY/F and YY/F, which accounted for 10.19%, 2.55%, 33.76%, 45.22%, 5.73% and 2.55%, respectively. CONCLUSION: High prevalence of Pfcrt CVIET and Pfmdr1 86Y, 184F double mutations confirm high-level CQ resistance (CQR) and might suggest reduced susceptibility of Plasmodium falciparum isolates to AQ in Bioko, EG. It establishes fundamental data for detection of P. falciparum CQR with molecular markers and will promotes the surveillance level of drug resistance in Bioko, EG.