Department of Veterinary Medicine, University of Bari, Bari, Italy.
STMicroelectronics Srl, Agrate Brianza (MB), Italy.
J Clin Microbiol. 2024 Jun 12;62(6):e0010424. doi: 10.1128/jcm.00104-24. Epub 2024 Apr 25.
Leishmaniasis is a vector-borne disease caused by many spp. which infect humans and other mammalian hosts. is the main agent of canine leishmaniasis (CanL) whose diagnosis is usually confirmed by serological and molecular tests. This study aimed to evaluate the clinical and analytical sensitivities of a lab-on-chip (LOC) real-time PCR applied on the portable Q3-Plus V2 platform (Q3 qPCR) in the detection of . The Q3 qPCR performance was assessed by comparing the quantification cycle (Cq) values with those obtained using the qPCR run on a CFX96 Real-Time System (CFX96 qPCR). A total of 173 DNA samples (extracted from bone marrow, lymph node, blood, buffy coat, conjunctival swab, and skin) as well as 93 non-extracted samples (NES) (bone marrow, lymph node, blood, and buffy coat) collected from dogs were tested with both systems. Serial dilutions of each representative DNA and NES sample were used to assess the analytical sensitivity of the Q3 qPCR system. Overlapping Cq values were obtained with the Q3 qPCR and CFX96 qPCR, both using DNA extracted from promastigotes (limit of detection, <1 promastigote per milliliter) and from biological samples as well as with NES. However, the Q3 qPCR system showed a higher sensitivity in detecting in NES as compared with the CFX96 qPCR. Our data indicate that the Q3 qPCR system could be a reliable tool for detecting DNA in biological samples, bypassing the DNA extraction step, which represents an advance in the point-of-care diagnostic of CanL.
利什曼病是一种由多种 spp. 引起的虫媒病,可感染人类和其他哺乳动物宿主。 是犬利什曼病(CanL)的主要病原体,其诊断通常通过血清学和分子检测来确认。本研究旨在评估应用于便携式 Q3-Plus V2 平台(Q3 qPCR)的芯片实验室(LOC)实时 PCR(qPCR)在检测 中的临床和分析灵敏度。通过比较定量循环(Cq)值与在 CFX96 实时系统(CFX96 qPCR)上运行的 qPCR 获得的 Cq 值来评估 Q3 qPCR 的性能。总共测试了 173 个 DNA 样本(从骨髓、淋巴结、血液、白细胞层、结膜拭子和皮肤中提取)和 93 个非提取样本(NES)(骨髓、淋巴结、血液和白细胞层),两个系统都进行了测试。使用每个代表性 DNA 和 NES 样本的系列稀释液来评估 Q3 qPCR 系统的分析灵敏度。使用从亲代虫(检测限,每毫升 <1 个亲代虫)和从生物样本中提取的 DNA 以及从 NES 中提取的 Q3 qPCR 和 CFX96 qPCR 均获得了重叠的 Cq 值。然而,与 CFX96 qPCR 相比,Q3 qPCR 系统在检测 NES 中的 时具有更高的灵敏度。我们的数据表明,Q3 qPCR 系统可能是一种可靠的工具,可用于检测生物样本中的 DNA,绕过 DNA 提取步骤,这是 CanL 即时诊断的一项进展。