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首页> 美国卫生研究院文献>Journal of Clinical Microbiology >Feasibility of Flow Cytometry for Measurements of Plasmodium falciparum Parasite Burden in Studies in Areas of Malaria Endemicity by Use of Bidimensional Assessment of YOYO-1 and Autofluorescence
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Feasibility of Flow Cytometry for Measurements of Plasmodium falciparum Parasite Burden in Studies in Areas of Malaria Endemicity by Use of Bidimensional Assessment of YOYO-1 and Autofluorescence

机译：利用YOYO-1的二维评估和自体荧光检测流式细胞术检测疟疾流行地区恶性疟原虫疟原虫负担的可行性。

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The detection and quantification of Plasmodium falciparum in studies of malaria endemicity primarily relies upon microscopy. High-throughput quantitative methods with less subjectivity and greater reliability are needed for investigational studies. The staining of parasitized erythrocytes with YOYO-1 for flow cytometry bears great potential as a tool for assessing malaria parasite burden. Capillary blood was collected from children presenting to the pediatric ward of the Manhiça District Hospital in Mozambique for parasitemia assessment by thick and thin blood films, flow cytometry (YOYO-1530/585), and quantitative real-time PCR (qRT-PCR). Whole blood was fixed and stained with YOYO-1 for acquisition on a cytometer to assess the frequency of infected erythrocyte events. qRT-PCR was used as the gold standard for the detection of P. falciparum. The YOYO-1530/585 method was as sensitive and specific as conventional microscopy (area under the receiver operating characteristic, 0.9 for both methods). The interrater mean difference for YOYO-1530/585 was near zero. Parasite density using flow cytometry and complete blood counts returned density estimates with a mean difference 2.2 times greater than results by microscopy (confidence interval, 1.46 to 3.60) but with limits of agreement between 10 times lower and 50 times higher than those of microscopy. The YOYO-1530/585 staining pattern was established exactly as demonstrated in animal models, but the assay was limited by the lack of appropriate negative-control samples for establishing background levels and the definition of positives in areas in which malaria is endemic. YOYO-1530/585 is a high-throughput tool with great potential if the limitations of negative controls and heterogeneous levels of background signal can be overcome.
机译：在疟疾流行性研究中，恶性疟原虫的检测和定量主要依靠显微镜。研究性研究需要主观性较低，可靠性更高的高通量定量方法。 YOYO-1染色的寄生红细胞用于流式细胞仪具有巨大的潜力，可作为评估疟疾寄生虫负担的工具。从出现在莫桑比克曼希卡地区医院儿科病房的儿童中收集毛细血管血，通过厚薄血膜，流式细胞仪（YOYO-1530 / 585）和定量实时PCR（qRT-PCR）评估寄生虫病。固定全血并用YOYO-1染色以在流式细胞仪上采集以评估感染的红细胞事件的频率。 qRT-PCR被用作检测恶性疟原虫的金标准。 YOYO-1530 / 585方法与常规显微镜一样灵敏和特异（在接收器工作特性下的面积，两种方法均为0.9）。 YOYO-1530 / 585的界面平均差接近零。使用流式细胞仪和全血细胞计数的寄生虫密度返回的密度估计值，其平均差值比显微镜下的结果大2.2倍（置信区间为1.46至3.60），但相差的极限是显微镜下的10倍至50倍。 YOYO-1530 / 585染色模式的建立与动物模型中所显示的完全一样，但是由于缺乏适当的阴性对照样品来建立背景水平以及疟疾流行地区的阳性定义，该测定法受到限制。 YOYO-1530 / 585是高通量的工具，如果可以克服阴性对照和背景信号的异类水平的局限性，则潜力巨大。

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