Development of a Digital Droplet Polymerase Chain Reaction (ddPCR) assay to detect Leishmania DNA in samples from Cutaneous Leishmaniasis patients.

Development of a Digital Droplet Polymerase Chain Reaction (ddPCR) assay to detect Leishmania DNA in samples from Cutaneous Leishmaniasis patients.
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DOI:
10.1016/j.ijid.2018.10.029
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发表时间:
2019-03
期刊:
International journal of infectious diseases : IJID : official publication of the International Society for Infectious Diseases
影响因子:
--
通讯作者:
Butcher R
Butcher R
中科院分区:
其他
文献类型:
--
作者:
Ramírez JD;Herrera G;Muskus C;Mendez C;Duque MC;Butcher R

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在这里,我们使用评估的qPCR评估ddPCR平台,方法以皮肤利什曼病患者的利什曼原虫培养物和临床标本为研究对象,建立了一种基于ddPCR的皮肤利什曼病诊断方法。结果该方法的检测限明显高于已发表的同类方法的检测限结论虽然ddPCR形式的该测定的性能对于研究目的是可接受的,但对于临床诊断目的是不够的。该检测试剂盒更适合qPCR平台。
AimHere, we evaluate the ddPCR platform using an evaluated qPCR-based diagnostic assay for the detection ofLeishmaniainfection in Cutaneous Leishmaniasis patients.MethodsA standard curve of culturedLeishmaniaparasite material and clinical samples of CL patients were tested with ddPCR to determine the analytical and diagnostic performance.ResultsThe limit of detection of the assay on the ddPCR platform was much higher than the published limit of detection of the same assay on the qPCR platform (100 vs 1 parasites/mL, respectively).ConclusionWhile the performance of this assay in ddPCR format was acceptable for research purposes, it is not sufficient for clinical diagnostic purposes. The assay is more suited to the qPCR platform.
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