Persistence of markers of chloroquine resistance among P. falciparum isolates recovered from two Nigerian communities.

MalariaWorld journal Pub Date : 2014-02-26 eCollection Date: 2014-01-01 DOI:10.5281/zenodo.10878545
Yetunde A Olukosi, Muyiwa K Oyebola, Olusola Ajibaye, Bassey A Orok, Olugbenga O Aina, Chimere O Agomo, Bamidele A Iwalokun, Samuel K Akindele, Veronica N V Enya, Hilary I Okoh
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Abstract

Background: A recovery in chloroquine efficacy following a period of cessation has raised the possibility of its reintroduction for malaria chemotherapy. We investigated the prevalence of the major markers of chloroquine resistance years after the withdrawal of the drug in Nigeria.

Materials and methods: Finger prick blood samples were collected from participants presenting with symptoms of malaria in two selected health centres each representing Lekki and Ijede communities of Lagos, Nigeria. Thick and thin blood smears were prepared for microscopy and dry blood spots made from malaria-positive participants for parasite DNA extraction. The detection of mutations in the Plasmodium falciparum chloroquine resistance transporter (pfcrt) and P. falciparum multidrug resistance (pfmdr1) genes was performed by nested polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP).

Results: Of the 1527 blood samples that were confirmed by PCR to be P. falciparum positive, 412 and 344 were typed for the molecular detection of pfcrt and pfmdr1 gene mutations, respectively. The mutant alleles of pfcrt were present among 290 (70%) parasite carriers while the pfmdr1 mutant allele was found in 117 (34%) of the total population. There were higher distributions of the mutant alleles for the two loci in Ijede than in Lekki. The observed frequencies of pfcrt mutant alleles in the two parasite populations were in agreement with the expected frequencies predicted by Hardy-Weinberg. In comparing data with studies conducted between 2000 and 2002 in Ijede, we observed an increase in the prevalence of mutant type pfcrt against a marginal decline in the pfmdr1 mutant type.

Conclusion: The high frequencies of pfcrt mutation are suggestive of a persistent drug pressure and continuing inefficacy of chloroquine as an antimalarial drug.

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从尼日利亚两个社区分离的恶性疟原虫耐氯喹标记物的持续性。
背景:氯喹停药一段时间后疗效有所恢复,因此有可能重新引入氯喹用于疟疾化疗。我们调查了尼日利亚停药数年后氯喹耐药性主要标志物的流行情况:在代表尼日利亚拉各斯莱基和伊杰德社区的两个选定医疗中心,分别从出现疟疾症状的患者身上采集手指刺血样本。制备了用于显微镜检查的厚血涂片和薄血涂片,并从疟疾阳性患者身上提取干血斑用于提取寄生虫 DNA。通过巢式聚合酶链反应(PCR)和限制性片段长度多态性(RFLP)检测恶性疟原虫氯喹抗性转运体(pfcrt)和恶性疟原虫多药抗性(pfmdr1)基因的突变:结果:经聚合酶链反应确认为恶性疟原虫阳性的1527份血样中,412份和344份分别进行了pfcrt和pfmdr1基因突变的分子检测。在 290 名(70%)寄生虫携带者中发现了 pfcrt 突变等位基因,而在 117 名(34%)携带者中发现了 pfmdr1 突变等位基因。这两个位点的突变等位基因在伊杰德的分布高于莱基。在两个寄生虫种群中观察到的 pfcrt 突变等位基因频率与 Hardy-Weinberg 预测的预期频率一致。与 2000 年至 2002 年期间在伊杰德进行的研究数据相比,我们发现 pfcrt 突变型的流行率有所上升,而 pfmdr1 突变型的流行率则略有下降:结论:pfcrt 突变的高频率表明,氯喹作为抗疟药物的药物压力持续存在,且效果不佳。
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