Mosquill Muzaffar, Nordin Syafinaz Amin, Ali Mohamad Ridhuan Mohd, Joseph Narcisse Mary Sither
Department of Medical Microbiology, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Selangor 43400, Malaysia.
Ministry of Health, Institute for Medical Research, Malaysia.
MethodsX. 2022 Jul 30;9:101804. doi: 10.1016/j.mex.2022.101804. eCollection 2022.
Leptospirosis is a zoonotic disease mostly occurring in tropical climate countries. The etiology of the disease is due to microbes from the genus . Higher number of cases reported worldwide indicated the disease is not easily eradicated. Leptospirosis shares the most common febrile symptoms such as dengue, Zika and yellow fever thus making it difficult to differentiate the disease at an early stage. The widely used current detection via PCR, uses the bacterial outer membrane protein (OMP) as their target region. However, the heterogeneity and variation of the genome cause false negative results. Lipoprotein LipL41 is the third most abundant outer membrane lipoprotein among pathogenic species and it is surface exposed and expressed during infection thus making it a suitable candidate in identifying pathogenic . LipL21 on the other hand is a potential candidate in identifying the intermediate species. The study aimed in designing suitable PCR primers in identifying pathogenic and intermediate species of through bioinformatics analysis on the bacterial OMPs. LipL41 and LipL21 were chosen as the suitable target sequence to be used as PCR primers in detecting the pathogenic and intermediate species, respectively. The designed primers indicated positive feedback upon tested with their respective bacterial DNA extract. These lipoproteins may serve as potential PCR primers to be used with clinical samples in diagnosing leptospirosis.•The etiology of the illness is due to bacteria from the genus .•PCR utilizes the bacterial external membrane protein (OMP) thus the heterogeneity and variety of the genome cause bogus adverse outcomes.•The suitable candidates are LipL41, the third most abundant outer membrane lipoprotein, whereas LipL21 is a potential candidate in identifying the intermediate species.
钩端螺旋体病是一种主要发生在热带气候国家的人畜共患病。该疾病的病因是由 属的微生物引起的。全球报告的病例数较多表明该疾病不易根除。钩端螺旋体病具有与登革热、寨卡病毒病和黄热病等最常见的发热症状相同，因此在早期难以区分该疾病。目前广泛使用的通过聚合酶链反应（PCR）进行的检测，将细菌外膜蛋白（OMP）作为其靶区域。然而，基因组的异质性和变异性会导致假阴性结果。脂蛋白LipL41是致病物种中第三丰富的外膜脂蛋白，它在感染期间暴露于表面并表达，因此使其成为鉴定病原体的合适候选物。另一方面，LipL21是鉴定中间物种的潜在候选物。该研究旨在通过对细菌OMPs进行生物信息学分析，设计合适的PCR引物来鉴定 的致病和中间物种。分别选择LipL41和LipL21作为合适的靶序列，用作检测致病和中间物种的PCR引物。所设计的引物在用各自的细菌DNA提取物进行测试时显示出阳性反馈。这些脂蛋白可作为潜在的PCR引物，用于临床样本中钩端螺旋体病的诊断。
•该疾病的病因是由 属的细菌引起的。
•PCR利用细菌外膜蛋白（OMP），因此基因组的异质性和多样性会导致假阴性结果。
•合适的候选物是LipL41，即第三丰富的外膜脂蛋白，而LipL21是鉴定中间物种的潜在候选物。
