Analysis of gene mutations involved in chloroquine resistance inPlasmodium falciparumparasites isolated from patients in the southwest of Saudi Arabia

Abstract

BACKGROUND AND OBJECTIVES:Chloroquine has been the drug of choice for the treatment of malaria for many decades. We aimed to examine the molecular basis of chloroquine resistance among Plasmodium falciparum isolates from the southwestern region of Saudi Arabia by analyzing the K76T and N86Y mutations in the PfCRT and PfMDR1 genes, respectively.PATIENTS AND METHODS:P falciparum-infected blood spot samples (n=121) were collected on filter papers. DNA was extracted and fragments from the above genes were amplified using nested PCR. The amplicons were digested by ApoI enzyme and sequenced.RESULTS:Of the 121 samples, 95 and 112 samples could be amplified for PfCRT K76T and PfMDR1 N86Y mutations, respectively. All of the samples amplified for the PfCRT K76T mutation were undigestible by ApoI, suggesting the presence of the K76T mutation. For the PfMDR1 N86Y mutation, 65/109 samples (59.6%) were digestible when treated with ApoI in a pattern, suggestive of the presence of the investigated wild allele (N86). However, 44/109 samples (40.4%) were digestible by ApoI, suggesting the presence of the mutated allele (Y) at position 86. DNA sequencing confirmed these results.CONCLUSION:Surprisingly, all isolates exhibited the mutated allele at codon 76 (K76T) of PfCRT. However, the mutated mutant allele at codon 86 (N86Y) of PfMDR1 was found in 40.4% of the samples studied. To our knowledge, this is the first study that has investigated the existence of the mutation in the PfMDR1 gene in the country. This study will contribute to the development of new strategies for therapeutic intervention against malaria in Saudi Arabia.

Genetic diversity of Plasmodium falciparum and distribution of drug resistance haplotypes in Yemen

“…The emergence of CQR in Yemen in 1989 Yemen [2,7,9,38,39], coincided with its appearance in close countries in East Africa [12,16,40] and Asia, Saudi Arabia [3], Iran and Pakistan [3,41-46]. The presence of two distinct pfcrt genotype, CV IET (89%) and S VMN T (4%) suggests that CQR in Yemen has evolved from at least two different origins.…”

“…Bahrain, Kuwait and United Arab Emirates (UAE) [1]. However, some sites in Yemen and southern Saudi Arabia remain malarious, with a high prevalence of drug-resistant Plasmodium falciparum parasites [2,3]. In addition, imported malaria cases, via asymptomatic travellers from malaria-endemic areas, sustain a major challenge for possible rebound of local transmission [4].…”

Molecular investigation of mixed malaria infections in Southwest Saudi Arabia

“…vivax is resistant to chloroquine, an appropriate ACT regimen is recommended together with a primaquine regimen. 4 Severe and complicated malaria is most commonly caused by P. falciparum , although P. vivax , and P. knowlesi may also be responsible for severe infections. Research on the P. vivax parasite has tended to lag behind, since it was previously thought that mono-infection with P. vivax resulted only in benign tertian fever, and that severe cases of P. vivax infection were the result of co-infection with P. falciparum .…”

Polymorphism of pfcrt K76T and pfatpase6 S769N Genes in Malaria Patients at Teluk Bintuni Regency, Papua, Indonesia

“…The pfcrt K76T gene was the gene associated with chloroquine resistance. 15,16 Present study suggested that the high level of chloroquine resistance in Teluk Bintuni area was found by the percentage of 81% samples showing pfcrt K76T gene mutation. This study was supported by the epidemiological study of malaria-endemic areas conducted by Syafruddin et al, which reported a positive correlation between pfcrt K76T gene polymorphism and chloroquine resistance with the polymorphism rate of 100% in western Indonesia (Nias, Hanura, Kokap and Kutai) and 91% in eastern Indonesia (Armopa, Flores, Mamuju and Minahasa).…”