Ibarra-Meneses Ana Victoria, Chicharro Carmen, Sánchez Carmen, García Emilia, Ortega Sheila, Ndung'u Joseph Mathu, Moreno Javier, Cruz Israel, Carrillo Eugenia
WHO Collaborating Centre for Leishmaniasis, National Center for Microbiology, Instituto de Salud Carlos III, 28220 Majadahonda, Spain.
Foundation for Innovative New Diagnostics, 1202 Geneva, Switzerland.
Microorganisms. 2021 Mar 16;9(3):610. doi: 10.3390/microorganisms9030610.
Loop-mediated isothermal amplification allows the rapid, sensitive and specific amplification of DNA without complex and expensive equipment. We compared the diagnostic performance of Loopamp™ Detection Kit (Eiken Chemical Co., Ltd., Tokyo, Japan) with conventional and real-time polymerase chain reaction (PCR) for human cutaneous and visceral leishmaniasis caused by . A total of 230 DNA samples from cutaneous (CL) and visceral (VL) leishmaniasis cases and controls from Spain, characterized by nested PCR (LnPCR) were tested by: (i) the Loopamp™ Detection Kit (Loopamp), run on Genie III real-time fluorimeter (OptiGene, UK); and (ii) real-time quantitative PCR (qPCR). The Loopamp test returned 98.8% (95% confidence interval-CI: 96.0-100.00) sensitivity and specificity of 97.7% (95% CI: 92.2-100) on VL samples, and 100% (95% CI: 99.1-100) sensitivity and 100.0% (95% CI: 98.8-100.0) specificity on CL samples. The Loopamp time-to-positivity () obtained by real-time fluorimetry showed excellent concordance (C = 97.91%) and strong correlation (r = 0.799) with qPCR's cycle threshold (). The performance of Loopamp is comparable to that of LnPCR and qPCR in the diagnosis of cutaneous and visceral leishmaniasis due to . The excellent correlation between the and should be further investigated to determine the accuracy of Loopamp to quantify parasite load in tissues.
环介导等温扩增技术可在无需复杂且昂贵设备的情况下实现DNA的快速、灵敏且特异的扩增。我们将环介导等温扩增检测试剂盒（Loopamp™ 检测试剂盒，日本东京荣研化学株式会社）与传统聚合酶链反应（PCR）及实时聚合酶链反应（PCR）针对由[具体病原体]引起的人类皮肤利什曼病和内脏利什曼病的诊断性能进行了比较。总共对230份来自西班牙皮肤利什曼病（CL）和内脏利什曼病（VL）病例及对照的DNA样本进行检测，这些样本经巢式PCR（LnPCR）鉴定，检测方法如下：（i）使用在英国OptiGene公司的Genie III实时荧光定量仪上运行的环介导等温扩增检测试剂盒（Loopamp）；（ii）实时定量PCR（qPCR）。环介导等温扩增检测在VL样本上的敏感性为98.8%（95%置信区间 - CI：96.0 - 100.00），特异性为97.7%（95% CI：92.2 - 100）；在CL样本上的敏感性为100%（95% CI：99.1 - 100），特异性为100.0%（95% CI：98.8 - 100.0）。通过实时荧光法获得的环介导等温扩增检测的阳性时间（）与qPCR的循环阈值（）显示出极好的一致性（C = 97.91%）和强相关性（r = 0.799）。在诊断由[具体病原体]引起的皮肤和内脏利什曼病方面，环介导等温扩增检测的性能与巢式PCR和qPCR相当。应进一步研究和之间的良好相关性，以确定环介导等温扩增检测在定量组织中寄生虫载量方面的准确性。
