Multiplex real-time PCR for the diagnosis of malaria: correlation with microscopy
Multiplex real-time PCR for the diagnosis of malaria: correlation with microscopy
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DOI：
10.1111/j.1469-0691.2010.03218.x
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发表时间：
2011-03-01
影响因子：
14.2
通讯作者：
Greub, G.
中科院分区：
文献类型：
--
作者：
Dormond, L.;Jaton-Ogay, K.;Greub, G.
疟疾通常通过显微镜和快速抗原检测来诊断。分子生物学方法得到了越来越广泛的应用。在本研究中，一个定量的多重疟疾PCR的贡献进行了调查。我们评估：（i）基于PCR的鉴定和显微镜检查之间的一致性，以及（ii）通过定量PCR和显微镜检查确定的寄生虫负荷之间的相关性。对于83例经显微镜检查为疟原虫属阳性的患者，通过多重PCR测试第一个EDTA-血液样品以确认基于涂片的物种鉴定。每天使用显微镜和PCR评估寄生虫负荷。在83例受试者中，1例仅镜检阳性，82例镜检和PCR均阳性。在种水平上，显微镜和PCR鉴定的一致性为89%（73/82）。9个不一致结果中有6个对应于两个或三个种属的合并感染，并归因于混合病例的形态学鉴定不准确。寄生虫负荷一般迅速下降后，治疗已经开始，使用显微镜和PCR获得类似的衰减曲线。我们的PCR证明特别有用的识别混合感染。通过PCR获得的定量与基于显微镜的定量密切相关，至少在临床试验中可用于监测治疗效果。
P>Malaria is generally diagnosed by microscopy and rapid antigen testing. Molecular methods become more widely used. In the present study, the contribution of a quantitative multiplex malaria PCR was investigated. We assessed: (i) the agreement between PCR-based identification and microscopy and (ii) the correlation between the parasite load as determined by quantitative PCR and by microscopy. For 83 patients positive by microscopy for Plasmodium spp., the first EDTA-blood sample was tested by multiplex PCR to confirm smear-based species identification. Parasite load was assessed daily using both microscopy and PCR. Among the 83 patients tested, one was positive by microscopy only and 82 were positive by microscopy and PCR. Agreement between microscopy and PCR for the identification at the species level was 89% (73/82). Six of the nine discordant results corresponded to co-infections by two or three species and were attributed to inaccurate morphological identification of mixed cases. The parasite load generally decreased rapidly after treatment had been started, with similar decay curves being obtained using both microscopy and PCR. Our PCR proved especially useful for identifying mixed infections. The quantification obtained by PCR closely correlated with microscopy-based quantification and could be useful for monitoring treatment efficacy, at least in clinical trials.