Submicroscopic malaria parasite carriage: how reproducible are polymerase chain reaction-based methods?
Submicroscopic malaria parasite carriage: how reproducible are polymerase chain reaction-based methods?
复制标题
亚显微疟疾寄生虫携带：基于聚合酶链反应的方法的重复性如何？
DOI：
10.1590/0074-0276140102
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发表时间：
2014-02
影响因子：
2.8
通讯作者：
Carvalho LH
中科院分区：
文献类型：
--
作者：
Costa DC;Madureira AP;Amaral LC;Sanchez BA;Gomes LT;Fontes CJ;Limongi JE;Brito CF;Carvalho LH
基于聚合酶链反应（PCR）的疟疾感染诊断方法有望准确识别亚显微寄生虫携带者。虽然已经描述了大量的PCR方案，但很少有研究讨论了在具有亚显微疟疾感染的现场样品的情况下PCR扩增的性能。在这里，两个完善的PCR方案（巢式PCR和实时PCR的疟原虫18小亚基rRNA基因）的再现性进行了评估，在一组34个血田样本的个人，是潜在的水库疟疾感染，但疟疾的光学显微镜呈阴性。无论采用哪种PCR方案，都观察到PCR重复之间存在很大差异，导致38%（13/34）的样本出现阳性和阴性交替结果。这些结果与从显微镜检查阳性患者或未暴露个体中获得的结果完全不同;这些个体的诊断可以基于基于PCR方案的高重现性和特异性来确认。PCR扩增的局限性仅限于寄生虫血症水平极低的田间样本，因为DNA模板的滴定能够检测血液中< 3个寄生虫/微升。总之，在亚显微疟疾感染的情况下，常规PCR方案需要仔细解释，因为可能出现不一致和假阴性结果。
The polymerase chain reaction (PCR)-based methods for the diagnosis of malaria infection are expected to accurately identify submicroscopic parasite carriers. Although a significant number of PCR protocols have been described, few studies have addressed the performance of PCR amplification in cases of field samples with submicroscopic malaria infection. Here, the reproducibility of two well-established PCR protocols (nested-PCR and real-time PCR for the Plasmodium 18 small subunit rRNA gene) were evaluated in a panel of 34 blood field samples from individuals that are potential reservoirs of malaria infection, but were negative for malaria by optical microscopy. Regardless of the PCR protocol, a large variation between the PCR replicates was observed, leading to alternating positive and negative results in 38% (13 out of 34) of the samples. These findings were quite different from those obtained from the microscopy-positive patients or the unexposed individuals; the diagnosis of these individuals could be confirmed based on the high reproducibility and specificity of the PCR-based protocols. The limitation of PCR amplification was restricted to the field samples with very low levels of parasitaemia because titrations of the DNA templates were able to detect < 3 parasites/µL in the blood. In conclusion, conventional PCR protocols require careful interpretation in cases of submicroscopic malaria infection, as inconsistent and false-negative results can occur.
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DOI：
10.4269/ajtmh.1996.55.647
发表时间：
1996-12-01
影响因子：
3.3
作者：
Jelinek, T;Proll, S;Kilian, AHD
通讯作者：
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发表时间：
2005-07-01
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