De Doncker Simonne, Hutse Veronik, Abdellati Saïd, Rijal Suman, Singh Karki Bal Man, Decuypere Saskia, Jacquet Diane, Le Ray Dominique, Boelaert Marleen, Koirala Shekhar, Dujardin Jean-Claude
Laboratory of Molecular Parasitology, Prins Leopold Instituut voor Tropische Geneeskunde, Nationalestraat 155, B-2000 Antwerp, Belgium.
Trans R Soc Trop Med Hyg. 2005 Jan;99(1):25-31. doi: 10.1016/j.trstmh.2004.01.015.
The PCR-ELISA represents a promising advance for diagnosis of visceral leishmaniasis (VL) in blood samples. However, the method has been validated mostly with HIV-positive patients who are known to have high levels of parasitaemia. We developed a new PCR-ELISA assay for specific detection of Leishmania in patients' blood and validated it in Nepalese subjects with clinically suspected VL, almost all of whom were HIV-negative. For blood samples, PCR-ELISA was more sensitive (83.9%) than conventional PCR (73.2%), and demonstrated 100% and 87.2% specificity when using healthy controls who had never travelled to a VL-endemic area and controls from a VL-endemic area as references, respectively. We have demonstrated the ability of PCR-ELISA to detect parasites in blood of HIV-negative patients. The method could be used for epidemiological as well as clinical purposes, as it reduces the need for traumatic bone marrow sampling and risky spleen aspiration.
PCR-ELISA法在血液样本中内脏利什曼病（VL）的诊断方面是一项很有前景的进展。然而，该方法大多是在已知寄生虫血症水平较高的HIV阳性患者中得到验证的。我们开发了一种用于特异性检测患者血液中利什曼原虫的新型PCR-ELISA检测方法，并在临床疑似VL的尼泊尔受试者中进行了验证，这些受试者几乎都是HIV阴性。对于血液样本，PCR-ELISA法比传统PCR法更敏感（83.9%），分别以从未前往过VL流行地区的健康对照和来自VL流行地区的对照作为参考时，其特异性分别为100%和87.2%。我们已经证明了PCR-ELISA法检测HIV阴性患者血液中寄生虫的能力。该方法可用于流行病学及临床目的，因为它减少了创伤性骨髓采样和有风险的脾脏穿刺的需求。
