BACKGROUND:Recent studies have demonstrated the deletion of the histidine-rich protein 2 (PfHRP2) gene (pfhrp2) in field isolates of Plasmodium falciparum, which could result in false negative test results when PfHRP2-based rapid diagnostic tests (RDTs) are used for malaria diagnosis. Although primary diagnosis of malaria in Honduras is determined based on microscopy, RDTs may be useful in remote areas. In this study, it was investigated whether there are deletions of the pfhrp2, pfhrp3 and their respective flanking genes in 68 P. falciparum parasite isolates collected from the city of Puerto Lempira, Honduras. In addition, further investigation considered the possible correlation between parasite population structure and the distribution of these gene deletions by genotyping seven neutral microsatellites. METHODS:Sixty-eight samples used in this study, which were obtained from a previous chloroquine efficacy study, were utilized in the analysis. All samples were genotyped for pfhrp2, pfhrp3 and flanking genes by PCR. The samples were then genotyped for seven neutral microsatellites in order to determine the parasite population structure in Puerto Lempira at the time of sample collection. RESULTS:It was found that all samples were positive for pfhrp2 and its flanking genes on chromosome 8. However, only 50% of the samples were positive for pfhrp3 and its neighboring genes while the rest were either pfhrp3-negative only or had deleted a combination of pfhrp3 and its neighbouring genes on chromosome 13. Population structure analysis predicted that there are at least two distinct parasite population clusters in this sample population. It was also determined that a greater proportion of parasites with pfhrp3-(and flanking gene) deletions belonged to one cluster compared to the other. CONCLUSION:The findings indicate that the P. falciparum parasite population in the municipality of Puerto Lempira maintains the pfhrp2 gene and that PfHRP2-based RDTs could be considered for use in this region; however continued monitoring of parasite population will be useful to detect any parasites with deletions of pfhrp2.

译文

背景:最近的研究表明,恶性疟原虫田间分离株中富含组氨酸的蛋白质2(PfHRP2)基因(pfhrp2)缺失,当使用基于PfHRP2的快速诊断测试(RDT)进行假阴性测试时,可能会导致假阴性结果。疟疾诊断。尽管洪都拉斯的疟疾初步诊断是根据显微镜检查确定的,但RDT在偏远地区可能有用。在这项研究中,调查了从洪都拉斯波多黎各伦皮拉市收集的68株恶性疟原虫的分离物中是否存在pfhrp2,pfhrp3及其各自侧翼基因的缺失。此外,进一步的研究考虑到通过对7个中性微卫星进行基因分型,寄生虫种群结构与这些基因缺失的分布之间可能存在相关性。
方法:本研究中使用的68个样品是从先前的氯喹功效研究中获得的,用于分析。通过PCR对所有样品的pfhrp2,pfhrp3和侧翼基因进行基因分型。然后对样品中的七个中性微卫星进行基因分型,以便在采样时确定波多黎各伦皮拉的寄生虫种群结构。
结果:发现所有样本的pfhrp2及其侧翼基因在8号染色体上均为阳性。但是,只有50%的样本的pfhrp3及其邻近基因为阳性,而其余样本仅为pfhrp3阴性或已删除组合pfhrp3及其邻近基因在13号染色体上的分布。种群结构分析预测,该样本群体中至少存在两个不同的寄生虫群体。还确定了具有pfhrp3-(和侧翼基因)缺失的较大比例的寄生虫与另一个簇相比属于一个簇。
结论:研究结果表明,波多黎各伦皮拉市的恶性疟原虫寄生虫种群维持pfhrp2基因,并且可以考虑在该区域使用基于PfHRP2的RDT。但是,继续监测寄生虫种群将有助于检测出带有pfhrp2缺失的任何寄生虫。

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