da C Gonçalves-de-Albuquerque Suênia, Pessoa-e-Silva Rômulo, Trajano-Silva Lays A M, de Morais Rayana C S, Brandão-Filho Sinval P, de Paiva-Cavalcanti Milena
Department of Immunology, Aggeu Magalhães Research Center, Oswaldo Cruz Foundation, Av. Prof. Moraes Rego, s/n, Recife, Pernambuco, 50670-420, Brazil,
Mol Biotechnol. 2015 Apr;57(4):318-24. doi: 10.1007/s12033-014-9825-2.
Early detection of leishmaniases and prompt institution of treatment are paramount for individuals and communities affected by these diseases. To overcome the remaining limitations inherent to molecular methods currently used and to ensure the accuracy of results in leishmaniases diagnosis, two triplex polymerase chain reaction (PCR) assays with quality controls for the reactions were developed. Validity indicators were assessed in 186 dog blood samples from endemic areas in Brazil. The level of agreement between the new tools and their singleplex protocols was assessed by kappa analysis. The triplex PCR for visceral leishmaniasis showed sensitivity (S) = 78.68 %, specificity (E) = 85.29 %, and efficiency (e) = 81.05 %. The cutaneous leishmaniasis protocol showed S = 97.29 %, E = 79.16 %, and e = 90.16 %. Both protocols showed good agreement with gold standards. These new tools enable, in a single reaction, the diagnosis of the diseases and the evaluation of the sample quality and DNA extraction process, thus reducing the cost of reagents and avoiding the eventual need for collecting a second sample.
对于受利什曼病影响的个人和社区而言，早期发现利什曼病并及时进行治疗至关重要。为克服目前使用的分子方法固有的剩余局限性，并确保利什曼病诊断结果的准确性，开发了两种带有反应质量控制的三重聚合酶链反应（PCR）检测方法。在来自巴西流行地区的186份犬血样本中评估了有效性指标。通过kappa分析评估了新工具与其单重检测方案之间的一致性水平。内脏利什曼病的三重PCR检测显示敏感性（S）=78.68%，特异性（E）=85.29%，效率（e）=81.05%。皮肤利什曼病检测方案显示S = 97.29%，E = 79.16%，e = 90.16%。两种检测方案均与金标准显示出良好的一致性。这些新工具能够在一次反应中诊断疾病并评估样本质量和DNA提取过程，从而降低试剂成本并避免最终需要采集第二个样本。
