Division of Bacteriology & Mycology, Indian Veterinary Research Institute, Izatnagar, 243122, India.
Division of Bacteriology & Mycology, Indian Veterinary Research Institute, Izatnagar, 243122, India.
Mol Cell Probes. 2017 Dec;36:29-35. doi: 10.1016/j.mcp.2017.08.001. Epub 2017 Aug 10.
Leptospirosis is considered to be the most widespread zoonotic disease caused by pathogenic species of Leptospira. The present study reports a novel set of primers targeting LigB gene for visual detection of pathogenic Leptospira in urine samples through Loop-mediated isothermal amplification (LAMP). The results were recorded by using Hydroxyl napthol blue (HNB), SYBR GREEN I and calcein. Analytical sensitivity of LAMP was as few as 10 leptospiral organisms in spiked urine samples from cattle and dog. LigB gene based LAMP, termed as LigB-LAMP, was found 10 times more sensitive than conventional PCR. The diagnostic specificity of LAMP was 100% when compared to SYBR green qPCR for detection of Leptospira in urine samples. Though qPCR was found more sensitive, the rapidity and simplicity in setting LAMP test followed by visual detection of Leptospira infection in clinical samples makes LigB-LAMP an alternative and favourable diagnostic tool in resource poor setting.
钩端螺旋体病被认为是最广泛的人畜共患疾病，由致病性钩端螺旋体引起。本研究报告了一套针对 LigB 基因的新引物，通过环介导等温扩增（LAMP）在尿液样本中对致病性钩端螺旋体进行可视化检测。结果通过羟基萘酚蓝（HNB）、SYBR GREEN I 和钙黄绿素进行记录。在来自牛和狗的尿液样本中，LAMP 的分析灵敏度低至 10 个钩端螺旋体生物。基于 LigB 基因的 LAMP 称为 LigB-LAMP，比常规 PCR 灵敏 10 倍。与用于检测尿液样本中钩端螺旋体的 SYBR 绿 qPCR 相比，LAMP 的诊断特异性为 100%。虽然 qPCR 被发现更灵敏，但 LAMP 试验的快速性和简单性以及随后对临床样本中钩端螺旋体感染的可视化检测使 LigB-LAMP 成为资源匮乏环境下的替代和有利的诊断工具。
