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柬埔寨低传播环境下疟疾的聚合酶链式反应检测:干血斑与静脉血样本对比

Malaria PCR detection in Cambodian low-transmission settings: dried blood spots versus venous blood samples.

作者信息

Canier Lydie, Khim Nimol, Kim Saorin, Eam Rotha, Khean Chanra, Loch Kaknika, Ken Malen, Pannus Pieter, Bosman Philippe, Stassijns Jorgen, Nackers Fabienne, Alipon SweetC, Char Meng Chuor, Chea Nguon, Etienne William, De Smet Martin, Kindermans Jean-Marie, Ménard Didier

机构信息

Institut Pasteur du Cambodge, Phnom Penh, Cambodia; Médecins Sans Frontières, Brussels, Belgium; Epicentre, Paris, France; Médecins Sans Frontières, Tuol Svay Prey I, Chamkarmon, Phnom Penh, Cambodia; National Center for Parasitology, Entomology and Malaria Control, Phnom Penh, Cambodia.

Institut Pasteur du Cambodge, Phnom Penh, Cambodia; Médecins Sans Frontières, Brussels, Belgium; Epicentre, Paris, France; Médecins Sans Frontières, Tuol Svay Prey I, Chamkarmon, Phnom Penh, Cambodia; National Center for Parasitology, Entomology and Malaria Control, Phnom Penh, Cambodia

出版信息

Am J Trop Med Hyg. 2015 Mar;92(3):573-7. doi: 10.4269/ajtmh.14-0614. Epub 2015 Jan 5.

Abstract

In the context of malaria elimination, novel strategies for detecting very low malaria parasite densities in asymptomatic individuals are needed. One of the major limitations of the malaria parasite detection methods is the volume of blood samples being analyzed. The objective of the study was to compare the diagnostic accuracy of a malaria polymerase chain reaction assay, from dried blood spots (DBS, 5 μL) and different volumes of venous blood (50 μL, 200 μL, and 1 mL). The limit of detection of the polymerase chain reaction assay, using calibrated Plasmodium falciparum blood dilutions, showed that venous blood samples (50 μL, 200 μL, 1 mL) combined with Qiagen extraction methods gave a similar threshold of 100 parasites/mL, ∼100-fold lower than 5 μL DBS/Instagene method. On a set of 521 field samples, collected in two different transmission areas in northern Cambodia, no significant difference in the proportion of parasite carriers, regardless of the methods used was found. The 5 μL DBS method missed 27% of the samples detected by the 1 mL venous blood method, but most of the missed parasites carriers were infected by Plasmodium vivax (84%). The remaining missed P. falciparum parasite carriers (N = 3) were only detected in high-transmission areas.

摘要

在疟疾消除的背景下,需要有新的策略来检测无症状个体中极低的疟原虫密度。疟原虫检测方法的主要局限性之一是所分析的血样体积。本研究的目的是比较疟原虫聚合酶链反应检测法对干血斑(DBS,5 μL)和不同体积静脉血(50 μL、200 μL和1 mL)的诊断准确性。使用校准的恶性疟原虫血液稀释液进行聚合酶链反应检测的检测限表明,静脉血样本(50 μL、200 μL、1 mL)与Qiagen提取方法相结合,得到的阈值相似,为每毫升100个疟原虫,比5 μL DBS/Instagene方法低约100倍。在柬埔寨北部两个不同传播地区收集的一组521份现场样本中,无论使用何种方法,疟原虫携带者的比例均未发现显著差异。5 μL DBS方法漏检了1 mL静脉血方法检测出的27%的样本,但大多数漏检的疟原虫携带者感染的是间日疟原虫(84%)。其余漏检的恶性疟原虫携带者(N = 3)仅在高传播地区被检测到。

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