Alizadeh Safar Ali, Abdolahpour Gholamreza, Pourmand Mohammad Reza, Naserpour Taghi, Najafipour Reza, Eshraghi Seyyed Saeed
Department of Pathobiology, School of Public Health and Institute of Public Health Research, Tehran University of Medical Sciences, Tehran, Iran.
Faculty of Veterinary Medicine, University of Tehran.
Iran J Microbiol. 2014 Jun;6(3):184-9.
Timely diagnosis of leptospirosis is essential for an effective treatment. Large diversity of clinical symptoms has led leptospirosis diagnosis difficult. Researchers have conducted many tests with wide-range of sensitivity and specificity to achieve novel diagnostic procedures which have higher sensitivity and specificity compared with previous tests and which are more reliable and available to public laboratories. This study aimed to introduce Lsa63 and LipL32 proteins-based ELISA tests with more sensitivity, specificity, accuracy and convenience for public laboratories.
Recombinant forms of Lsa63 and LipL32 proteins were first generated. After coating these proteins, IgM and IgG ELISA tests were performed. 220 patients with suspicion of leptospirosis infection were selected for serum collection. The sera tests were carried out using MAT, IgM and IgG ELISA tests. In order to assess the performance of ELISA, the results of this test were compared with MAT.
30% of serum samples (n=65) in MAT were positive for leptospirosis infection, while ELISA tests including rLipL32- rLsa63-IgM and rLipL32-rLsa63-IgG showed 40.45% (n=89) and 38.63% (n=80) positive reaction, respectively.
Our results demonstrated that new ELISA tests based on mixing LipL32 and Lsa63 proteins, a novel mixture of recombinant antigens, are valuable to detect specific antibodies against pathogenic Leptospira in human serum and could be considered as helpful techniques in leptospirosis diagnosis.
钩端螺旋体病的及时诊断对于有效治疗至关重要。临床症状的高度多样性导致钩端螺旋体病诊断困难。研究人员进行了许多具有广泛敏感性和特异性的检测，以实现新的诊断程序，这些程序与以前的检测相比具有更高的敏感性和特异性，并且对公共实验室更可靠且可用。本研究旨在为公共实验室引入基于Lsa63和LipL32蛋白的ELISA检测，其具有更高的敏感性、特异性、准确性和便利性。
首先制备Lsa63和LipL32蛋白的重组形式。包被这些蛋白后，进行IgM和IgG ELISA检测。选择220例疑似钩端螺旋体感染的患者进行血清采集。使用显微镜凝集试验（MAT）、IgM和IgG ELISA检测进行血清检测。为了评估ELISA的性能，将该检测结果与MAT结果进行比较。
MAT检测中30%的血清样本（n = 65）钩端螺旋体感染呈阳性，而包括rLipL32 - rLsa63 - IgM和rLipL32 - rLsa63 - IgG的ELISA检测分别显示40.45%（n = 89）和38.63%（n = 80）呈阳性反应。
我们的结果表明，基于混合LipL32和Lsa63蛋白（一种新型重组抗原混合物）的新型ELISA检测对于检测人血清中针对致病性钩端螺旋体的特异性抗体具有重要价值，可被视为钩端螺旋体病诊断中的有用技术。
