caa a22 4500 445341491 CHVBK 20180317142814.0 cr unu---uuuuu 170323e20110801xx s 000 0 eng 10.1007/s00436-011-2251-5 doi (NATIONALLICENCE)springer-10.1007/s00436-011-2251-5 Detecting mutations in PfCRT and PfMDR1 genes among Plasmodium falciparum isolates from Saudi Arabia by pyrosequencing [Elektronische Daten] [Saad Bin Dajem, Adel Al-Sheikh, Marie Bohol, Mohammad Alhawi, Mohammed Al-Ahdal, Ahmed Al-Qahtani] The emergence of chloroquine resistance in Plasmodium falciparum is a significant public health problem where malaria is endemic. We aimed to evaluate the efficacy of pyrosequencing to assess chloroquine resistance among P. falciparum isolates from the southwestern region of Saudi Arabia by analyzing the K76T and N86Y mutations in the P. falciparum chloroquine resistance transporter (PfCRT) and P. falciparum multidrug resistance 1 (PfMDR1) genes, respectively. Blood samples (n = 121) from microscopically positive P. falciparum cases were collected. DNA was extracted, and fragments from each of the genes were amplified by PCR using new sets of primers. The amplicons were sequenced using a pyrosequencer. All of the 121 samples were amplified for assessment of the PfCRT K76T and PfMDR1 N86Y mutations. All of the samples amplified for the PfCRT 76T mutation harbored the ACA codon (121/121; 100%), indicating the presence of the 76T mutation. For the PfMDR1 N86Y mutation, 72/121 samples (59.5%) had the sequence AAT at that position, indicating the presence of the wild-type allele (86N). However, 49/121 samples (40.5%) had a TAT codon, indicating the mutant allele (Y) at position 86. This study shows that pyrosequencing could be useful as a high throughput, rapid, and sensitive assay for the detection of specific single nucleotide polymorphisms in drug-resistant P. falciparum strains. This will help health authorities in malaria-endemic regions to adopt new malaria control strategies that will be applicable for diagnostic and drug resistance assays for malaria and other life-threatening pathogens that are endemic in their respective countries. Springer-Verlag, 2011 Bin Dajem Saad Biology Department, College of Science, King Khalid University, Abha, Saudi Arabia aut Al-Sheikh Adel National Center for Training and Research, Ministry of Health, Jazan, Saudi Arabia aut Bohol Marie Department of Biological and Medical Research, King Faisal Specialist Hospital and Research Center, MBC 03, PO Box3354, 11211, Riyadh, Saudi Arabia aut Alhawi Mohammad College of Health Sciences, King Abdulaziz University, Jeddah, Saudi Arabia aut Al-Ahdal Mohammed Department of Biological and Medical Research, King Faisal Specialist Hospital and Research Center, MBC 03, PO Box3354, 11211, Riyadh, Saudi Arabia aut Al-Qahtani Ahmed Department of Biological and Medical Research, King Faisal Specialist Hospital and Research Center, MBC 03, PO Box3354, 11211, Riyadh, Saudi Arabia aut Parasitology Research Springer-Verlag 109/2(2011-08-01), 291-296 0932-0113 109:2<291 2011 109 436 https://doi.org/10.1007/s00436-011-2251-5 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s00436-011-2251-5 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Bin Dajem Saad Biology Department, College of Science, King Khalid University, Abha, Saudi Arabia aut NATIONALLICENCE 700 1- Al-Sheikh Adel National Center for Training and Research, Ministry of Health, Jazan, Saudi Arabia aut NATIONALLICENCE 700 1- Bohol Marie Department of Biological and Medical Research, King Faisal Specialist Hospital and Research Center, MBC 03, PO Box3354, 11211, Riyadh, Saudi Arabia aut NATIONALLICENCE 700 1- Alhawi Mohammad College of Health Sciences, King Abdulaziz University, Jeddah, Saudi Arabia aut NATIONALLICENCE 700 1- Al-Ahdal Mohammed Department of Biological and Medical Research, King Faisal Specialist Hospital and Research Center, MBC 03, PO Box3354, 11211, Riyadh, Saudi Arabia aut NATIONALLICENCE 700 1- Al-Qahtani Ahmed Department of Biological and Medical Research, King Faisal Specialist Hospital and Research Center, MBC 03, PO Box3354, 11211, Riyadh, Saudi Arabia aut NATIONALLICENCE 773 0- Parasitology Research Springer-Verlag 109/2(2011-08-01), 291-296 0932-0113 109:2<291 2011 109 436 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer