Secretaria de Estado da Saúde de São Paulo, São Paulo, SP, Brasil.
Mem Inst Oswaldo Cruz. 2011 Sep;106(6):691-700. doi: 10.1590/s0074-02762011000600008.
Malaria diagnoses has traditionally been made using thick blood smears, but more sensitive and faster techniques are required to process large numbers of samples in clinical and epidemiological studies and in blood donor screening. Here, we evaluated molecular and serological tools to build a screening platform for pooled samples aimed at reducing both the time and the cost of these diagnoses. Positive and negative samples were analysed in individual and pooled experiments using real-time polymerase chain reaction (PCR), nested PCR and an immunochromatographic test. For the individual tests, 46/49 samples were positive by real-time PCR, 46/49 were positive by nested PCR and 32/46 were positive by immunochromatographic test. For the assays performed using pooled samples, 13/15 samples were positive by real-time PCR and nested PCR and 11/15 were positive by immunochromatographic test. These molecular methods demonstrated sensitivity and specificity for both the individual and pooled samples. Due to the advantages of the real-time PCR, such as the fast processing and the closed system, this method should be indicated as the first choice for use in large-scale diagnosis and the nested PCR should be used for species differentiation. However, additional field isolates should be tested to confirm the results achieved using cultured parasites and the serological test should only be adopted as a complementary method for malaria diagnosis.
疟疾诊断传统上采用厚血涂片，但在临床和流行病学研究以及献血者筛查中，需要更敏感和更快的技术来处理大量样本。在这里，我们评估了分子和血清学工具，以建立一个用于混合样本的筛选平台，旨在减少这些诊断的时间和成本。使用实时聚合酶链反应（PCR）、巢式 PCR 和免疫层析试验对个体和混合实验中的阳性和阴性样本进行分析。对于个体测试，实时 PCR 检测出 46/49 份阳性样本，巢式 PCR 检测出 46/49 份阳性样本，免疫层析试验检测出 32/46 份阳性样本。对于使用混合样本进行的检测，实时 PCR 和巢式 PCR 检测出 13/15 份阳性样本，免疫层析试验检测出 11/15 份阳性样本。这些分子方法对个体和混合样本均具有敏感性和特异性。由于实时 PCR 具有快速处理和封闭系统等优点，因此该方法应被推荐用于大规模诊断，而巢式 PCR 应用于物种分化。然而，应测试额外的现场分离株以确认使用培养寄生虫获得的结果，并且应仅将血清学试验作为疟疾诊断的补充方法。
