Khatun Mahbuba, Alam S M Sabbir, Khan Abed Hussain, Hossain M Anwar, Haq Jalaluddin Ashraful, Alam Jilani Md Shariful, Rahman Mohammad Tariqur, Karim Muhammad Manjurul
Department of Microbiology, University of Dhaka, Dhaka 1000, Bangladesh.
Department of Medicine, Bangabandhu Sheikh Mujib Medical University, Dhaka 1000, Bangladesh.
Asian Pac J Trop Med. 2017 Aug;10(8):753-759. doi: 10.1016/j.apjtm.2017.08.002. Epub 2017 Aug 21.
OBJECTIVE: To establish a suitable method of diagnosis of visceral leishmaniasis (VL) using peripheral blood, spleen or bone marrow aspirates. METHODS: Peripheral blood, bone marrow and spleen aspirate samples were collected from clinically suspected VL patients (n = 26). A new PCR primer pair (MK1F/R) was designed targeting kinetoplast mini circle DNA sequences of Leishmania donovani, and Leishmania infantum, and was used to diagnose VL along with some other established primers for VL in polymerase chain reactions. Test was validated by comparing with several other diagnostic methods. RESULTS: The designed primer set showed 100% specificity and 98% sensitivity in detecting VL using blood samples, when compared with more invasive samples: bone marrow or spleen aspirates. CONCLUSIONS: The newly designed primer MK1F/R could be a better alternative for PCR based diagnosis of VL using less invasive sample, peripheral blood instead of bone marrow or spleen aspirates.
目的：建立一种利用外周血、脾脏或骨髓穿刺液诊断内脏利什曼病（VL）的合适方法。 方法：从临床疑似VL患者（n = 26）中采集外周血、骨髓和脾脏穿刺液样本。设计了一对新的PCR引物对（MK1F/R），靶向杜氏利什曼原虫和婴儿利什曼原虫的动基体小环DNA序列，并在聚合酶链反应中与其他一些已有的VL引物一起用于诊断VL。通过与其他几种诊断方法比较来验证该检测。 结果：与更具侵入性的样本（骨髓或脾脏穿刺液）相比，设计的引物组在使用血液样本检测VL时显示出100%的特异性和98%的敏感性。 结论：新设计的引物MK1F/R可以成为基于PCR诊断VL的更好选择，使用侵入性较小的样本——外周血而非骨髓或脾脏穿刺液。
Asian Pac J Trop Med. 2017-8
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