Medical Mycology and Bacteriology Research Center, Kerman University of Medical Sciences, Kerman, Iran.
Leishmaniasis Research Centre, Kerman University of Medical Sciences, Kerman, Iran.
Microb Pathog. 2022 Aug;169:105612. doi: 10.1016/j.micpath.2022.105612. Epub 2022 May 31.
Leishmaniasis is a neglected and widespread parasitic disease that can lead to serious health problems. The conventional method in diagnostic health clinics is direct smear preparation of the lesion and staining with standard Giemsa to visualize the amastigote stage and by culturing the organism in an NNN (Novy-MacNeal-Nicolle) to observe the promastigote form of the parasite. In the case of urban-type leishmaniasis, microscopic diagnosis is sometimes not possible due to the reduction of amastigotes in patients' wounds. Because most endemic areas are located in regions that do not have access to laboratories equipped with molecular tools, access to a rapid test to diagnose the disease is essential. In this study, for the first time for DNA extraction, the scalpel used for sampling was washed and extracted by boiling method. Also, the LAMP technique in this study was modified so that the test can be performed in 10 min and the results can be recognized by color. We used four microscopic methods, conventional PCR, real-time PCR, and LAMP, to diagnose urban-type leishmaniasis and compared the results of these methods with each other. The sensitivity and specificity of LAMP were higher than other techniques used. Therefore, it allows rapid diagnosis for timely treatment of the disease to control the primary reservoir host more quickly in ACL as humans are the principal source of infection. This test is performed at a high-speed and is cost-effective. For its convenience, this test is highly recommended to be used in endemic areas.
利什曼病是一种被忽视的广泛寄生虫病，可导致严重的健康问题。诊断保健诊所的常规方法是对病变进行直接涂片准备，并使用标准吉姆萨染色来观察无鞭毛体阶段，并通过在 NNN（Novy-MacNeal-Nicolle）中培养该生物体来观察寄生虫的前鞭毛体形式。在城市型利什曼病的情况下，由于患者伤口中无鞭毛体的减少，有时无法进行显微镜诊断。由于大多数流行地区位于无法获得配备分子工具的实验室的地区，因此获得用于诊断该疾病的快速测试至关重要。在这项研究中，首次使用手术刀进行采样，并通过煮沸法清洗和提取 DNA。此外，本研究还对 LAMP 技术进行了修改，使测试可以在 10 分钟内完成，并且可以通过颜色识别结果。我们使用了四种显微镜方法，即常规 PCR、实时 PCR 和 LAMP，来诊断城市型利什曼病，并相互比较了这些方法的结果。LAMP 的灵敏度和特异性高于其他使用的技术。因此，它可以快速诊断，以便及时治疗疾病，从而在 ACL 中更快地控制主要储存宿主，因为人类是感染的主要来源。该测试可高速进行，并且具有成本效益。出于方便考虑，强烈建议在流行地区使用该测试。
