用于钩端螺旋体病检测的重组LipL32蛋白的获取与鉴定

Obtaining and characterization of a recombinant LipL32 protein for detection of leptospirosis.

作者信息

Zhylkibayev A, Akishev Z, Khassenov B, Sarina N, Ramankulov Y, Mukanov K, Eskendirova S

机构信息

National Center for Biotechnology, 13/5 Korgalzhyn Road, Astana, Kazakhstan, 010000.

Nazarbayev University, School of Science and Technology, 53 Kabanbai batyra Avenue, Astana, Kazakhstan, 010000.

出版信息

Trop Biomed. 2018 Mar 1;35(1):280-287.

PMID:33601801

Abstract

Leptospirosis is a disease which affects domestic and wild animals as well as humans. A highly conserved outer membrane lipoprotein LipL32 is considered to be an important antigen for detection of leptospirosis. In this study the recombinant protein LipL32 was extracted and characterized by immunochemical methods. The recombinant protein LipL32 was produced in Escherichia coli. We have optimized a LipL32 gene for expression in Rosetta (DE3) strain. 0.5% sodium lauryl sulfate was used to solubilize the protein and expressed into inclusion bodies. Immunochemical studies demonstrated a high reactivity of the recombinant LipL32 with antibodies elicited during infection of pathogenic Leptospira serovars. This study tested the diagnostic performance of the recombinant LipL32 in an in-house ELISA. The indirect ELISA was developed and its specificity was compared to that of a traditional microagglutination test (MAT).

摘要

钩端螺旋体病是一种影响家畜、野生动物以及人类的疾病。一种高度保守的外膜脂蛋白LipL32被认为是检测钩端螺旋体病的重要抗原。在本研究中，重组蛋白LipL32被提取出来并通过免疫化学方法进行表征。重组蛋白LipL32在大肠杆菌中产生。我们优化了LipL32基因以在Rosetta (DE3)菌株中表达。使用0.5%的十二烷基硫酸钠溶解该蛋白并使其表达形成包涵体。免疫化学研究表明重组LipL32与致病性钩端螺旋体血清型感染期间产生的抗体具有高反应性。本研究在内部酶联免疫吸附测定（ELISA）中测试了重组LipL32的诊断性能。开发了间接ELISA并将其特异性与传统的显微凝集试验（MAT）进行比较。