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首页> 外文期刊>Parasitology >Development and validation of flow cytometric measurement for parasitaemia using autofluorescence and YOYO-1 in rodent malaria
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Development and validation of flow cytometric measurement for parasitaemia using autofluorescence and YOYO-1 in rodent malaria

机译：使用自体荧光和YOYO-1检测啮齿类疟疾中寄生虫血症的流式细胞仪的开发和验证

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An automated flow cytometric (FCM) detection method has been developed and validated with a simple diagnostic procedure in parasitized erythrocytes of Plasmodium berghei-infected rats using the nucleic acid-binding fluorescent dye YOYO-1. High levels of reticulocytes were detected during the course of the infection, ranging from 1.2-51.2%, but any RNA potentially confounding the assay could be removed by digestion with RNAse. The cell counts of uninfected, infected, and nucleated cells occurred with high precision. The cells were divided into different populations according to their physical or chemical properties but various factors within the assay such as cell fixation, RNA digestion, and compensation required optimization. In this study, FCM greatly simplified and accelerated parasite detection, with a mean precision of 4.4%, specificity of 98.9% and accuracy of 101.3%. The detection and quantitation limits in the assay were 0.024% and 0.074% parasitaemia, respectively. Overall, the parasite countsby FCM measurement correlated highly (r2=0.954-0. 988) with the parasitaemia measured by light microscopical analysis when animals treated with suppressive, clearance, and curative doses of novel antimalarial drugs were examined. The lower levels of parasitaemia (30%) detected by microscopy compared to FCM may be related to a number of schizonts externally attached to the erythrocyte membranes that normally would not be included in microscopy counting. Lower sampling error and reliable identification of rodent erythrocyte parasites based on the principles of FCM have replaced the traditional blood smear in our laboratory. 2007 Cambridge University Press.
机译：已经开发了一种自动流式细胞术（FCM）检测方法，并使用核酸结合荧光染料YOYO-1在简单的诊断程序中验证了伯氏疟原虫感染大鼠的寄生红细胞。在感染过程中检测到高水平的网织红细胞，范围为1.2-51.2％，但是任何可能混淆测定的RNA都可以通过用RNAse消化来去除。未感染，已感染和有核细胞的细胞计数发生率很高。根据细胞的物理或化学性质将其分为不同的种群，但测定中的各种因素（例如细胞固定，RNA消化和补偿）需要优化。在这项研究中，FCM大大简化并加速了寄生虫检测，平均准确度为4.4％，特异性为98.9％，准确度为101.3％。测定中的检出限和定量限分别为寄生虫血症的0.024％和0.074％。总体而言，当检查接受抑制，清除和治愈剂量的新型抗疟疾药物治疗的动物时，通过FCM测量得出的寄生虫计数与通过光学显微镜分析测得的寄生虫血症高度相关（r2 = 0.954-0.988）。与FCM相比，通过显微镜检查发现的寄生虫血症水平较低（30％），可能与外部附着在红细胞膜上的裂殖体的数量有关，这些裂殖体通常不会包括在显微镜计数中。更低的采样误差和基于FCM原理的啮齿动物红细胞寄生虫的可靠识别已取代了我们实验室中的传统血液涂片检查。 2007剑桥大学出版社。

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