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首页> 外文期刊>Malaria Journal >Sub-microscopic malaria cases and mixed malaria infection in a remote area of high malaria endemicity in Rattanakiri province, Cambodia: implication for malaria elimination
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Sub-microscopic malaria cases and mixed malaria infection in a remote area of high malaria endemicity in Rattanakiri province, Cambodia: implication for malaria elimination

机译：柬埔寨拉塔纳基里省偏远地区疟疾高发地区的亚微观疟疾病例和混合疟疾感染：消除疟疾的意义

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Background Malaria microscopy and rapid diagnostic tests are insensitive for very low-density parasitaemia. This insensitivity may lead to missed asymptomatic sub-microscopic parasitaemia, a potential reservoir for infection. Similarly, mixed infections and interactions between Plasmodium species may be missed. The objectives were first to develop a rapid and sensitive PCR-based diagnostic method to detect low parasitaemia and mixed infections, and then to investigate the epidemiological importance of sub-microscopic and mixed infections in Rattanakiri Province, Cambodia. Methods A new malaria diagnostic method, using restriction fragment length polymorphism analysis of the cytochrome b genes of the four human Plasmodium species and denaturing high performance liquid chromatography, has been developed. The results of this RFLP-dHPLC method have been compared to 1) traditional nested PCR amplification of the 18S rRNA gene, 2) sequencing of the amplified fragments of the cytochrome b gene and 3) microscopy. Blood spots on filter paper and Giemsa-stained blood thick smears collected in 2001 from 1,356 inhabitants of eight villages of Rattanakiri Province have been analysed by the RFLP-dHPLC method and microscopy to assess the prevalence of sub-microscopic and mixed infections. Results The sensitivity and specificity of the new RFLP-dHPLC was similar to that of the other molecular methods. The RFLP-dHPLC method was more sensitive and specific than microscopy, particularly for detecting low-level parasitaemia and mixed infections. In Rattanakiri Province, the prevalences of Plasmodium falciparum and Plasmodium vivax were approximately two-fold and three-fold higher, respectively, by RFLP-dHPLC (59% and 15%, respectively) than by microscopy (28% and 5%, respectively). In addition, Plasmodium ovale and Plasmodium malariae were never detected by microscopy, while they were detected by RFLP-dHPLC, in 11.2% and 1.3% of the blood samples, respectively. Moreover, the proportion of mixed infections detected by RFLP-dHPLC was higher (23%) than with microscopy (8%). Conclusions The rapid and sensitive molecular diagnosis method developed here could be considered for mass screening and ACT treatment of inhabitants of low-endemicity areas of Southeast Asia.
机译：背景疟疾显微镜检查和快速诊断检查对极低密度的寄生虫血症不敏感。这种不敏感可能会导致无症状的亚显微寄生性疟原虫遗漏，这是潜在的感染源。同样，疟原虫物种之间的混合感染和相互作用可能会丢失。目标首先是开发一种基于PCR的快速灵敏的诊断方法，以检测低寄生虫血症和混合感染，然后研究柬埔寨Rattanakiri省亚显微感染和混合感染的流行病学重要性。方法利用4种人类疟原虫细胞色素b基因的限制性片段长度多态性分析和变性高效液相色谱法，开发了一种新的疟疾诊断方法。将该RFLP-dHPLC方法的结果与1）18S rRNA基因的传统巢式PCR扩增，2）细胞色素b基因的扩增片段测序和3）显微镜相比较。通过RFLP-dHPLC方法和显微镜分析了2001年从Rattanakiri省的8个村庄的1356名居民中收集的滤纸上的血斑和Giemsa染色的血厚涂片，并通过RFLP-dHPLC方法进行了分析，以评估亚显微和混合感染的患病率。结果新型RFLP-dHPLC的灵敏度和特异性与其他分子方法相似。与显微镜相比，RFLP-dHPLC方法更灵敏，更具特异性，特别是对于检测低水平的寄生虫血症和混合感染。在拉塔纳基里省，通过RFLP-dHPLC（分别为59％和15％），恶性疟原虫和间日疟原虫的患病率分别比显微镜检查（分别为28％和5％）高2倍和3倍。 。此外，从未在显微镜下检测到卵圆形疟原虫和疟疾疟原虫，而通过RFLP-dHPLC分别在11.2％和1.3％的血液样本中检测到了它们。此外，RFLP-dHPLC检测到的混合感染比例（23％）高于显微镜检查（8％）。结论本文开发的快速灵敏的分子诊断方法可用于东南亚低流行地区居民的大规模筛查和ACT治疗。

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