Rahim Samiur, Sharif Md Mohiuddin, Amin Md Robed, Rahman Mohammad Tariqur, Karim Muhammad Manjurul
Department of Microbiology, University of Dhaka, Dhaka, Bangladesh.
Department of Medicine, Dhaka Medical College, Dhaka, Bangladesh.
PLOS Glob Public Health. 2022 Dec 29;2(12):e0000834. doi: 10.1371/journal.pgph.0000834. eCollection 2022.
Diagnosis of visceral leishmaniasis (VL) through the detection of its causative agents namely Leishmania donovani and L. infantum is traditionally based on immunochromatographic tests, microscopy of bone marrow, spleen aspirates, liver or lymph node and differential diagnosis. While the first process has low specificity, the later one carries the risk of fatal hemorrhage. Over the last decade, multiple Polymerase Chain Reaction (PCR) based diagnosis has been developed using blood and urine sample with a varying degree of sensitivity and specificity, an issue worth improving for precision diagnosis. Earlier, we reported a PCR-based diagnosis of L. donovani in peripheral blood using a novel set of PCR primers with absolute specificity. Using the same set of primers and PCR conditions, here we describe diagnosis of L. donovani from urine, for a non-invasive, rapid and safe diagnosis. Diagnosis of VL was carried out using urine samples collected from clinically diagnosed VL patients (n = 23) of Bangladesh in Real Time PCR. Test results were validated by comparing blood samples from the same set of patients. Sensitivity and specificity of this diagnosis was analyzed using retrospective bone marrow samples, collected earlier from confirmed VL patients (n = 19). The method showed 100% sensitivity in detecting L. donovani in urine and corresponding blood and retrospective bone marrow samples, as well as 100% specificity in control groups. A Real Time PCR-based molecular detection system using urine sample is hereafter presented what could be a, non-invasive approach for VL detection with precision and perfection.
内脏利什曼病（VL）的诊断传统上是通过检测其病原体，即杜氏利什曼原虫和婴儿利什曼原虫，基于免疫层析试验、骨髓显微镜检查、脾脏穿刺物、肝脏或淋巴结以及鉴别诊断。虽然第一种方法特异性较低，但后一种方法存在致命出血的风险。在过去十年中，已经开发了多种基于聚合酶链反应（PCR）的诊断方法，使用血液和尿液样本，其灵敏度和特异性各不相同，这是一个值得改进以实现精确诊断的问题。此前，我们报道了一种使用具有绝对特异性的新型PCR引物对外周血中的杜氏利什曼原虫进行基于PCR的诊断。使用相同的引物组和PCR条件，在此我们描述从尿液中诊断杜氏利什曼原虫，以实现无创、快速和安全的诊断。使用从孟加拉国临床诊断为VL的患者（n = 23）收集的尿液样本通过实时PCR进行VL诊断。通过比较同一组患者的血液样本对测试结果进行验证。使用早期从确诊的VL患者（n = 19）收集的回顾性骨髓样本分析该诊断方法的灵敏度和特异性。该方法在检测尿液以及相应血液和回顾性骨髓样本中的杜氏利什曼原虫时显示出100%的灵敏度，在对照组中也显示出100%的特异性。在此展示一种基于实时PCR的分子检测系统，该系统使用尿液样本，可能是一种精确完美的VL无创检测方法。
