Recombinant LipL32 Protein for Leptospirosis Detection in Indonesia

Sumarningsih, Simson Tarigan, Susanti, Kusmiyati
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引用次数: 8

Abstract

Leptospirosis is an endemic zoonotic disease in tropical countries. However, detection using the microagglutination test (MAT) as a gold standard is difficult to perform in Indonesia. Therefore, recombinant LipL32 protein (rLipL32) has been studied as an antigen for an ELISA test to detect Leptospirosis. We produced rLipL32 using the pRSET-C vector and expressed it in E. coli BL21 (DE3) competent cells. Under native conditions, we purified 2 mg rLipL32 protein from 500 ml culture. Analysis using western blotting and ELISA showed that serum from the bovine positive MAT serovar Hardjo could recognize pure rLipL32 protein. This result confirms an earlier study that indicates that rLipL32 protein is a good antigen for Leptospirosis detection. The diagnostic assay using rLipL32 is safe because it does not use infectious bacteria as an antigen and because it is easy to perform in every diagnostic laboratory in Indonesia. However, further study is still required for field validation of the rLipL32 assay.

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重组LipL32蛋白检测印尼钩端螺旋体病
钩端螺旋体病是热带国家的一种地方性人畜共患病。然而,在印度尼西亚很难将微凝集试验(MAT)作为金标准进行检测。因此,我们研究了重组LipL32蛋白(rLipL32)作为钩端螺旋体病ELISA检测的抗原。我们使用pRSET-C载体产生rLipL32,并在大肠杆菌BL21 (DE3)感态细胞中表达。在自然条件下,我们从500 ml培养物中纯化了2 mg rLipL32蛋白。western blotting和ELISA分析表明,牛MAT阳性血清Hardjo能识别纯rLipL32蛋白。这一结果证实了早期的研究表明rLipL32蛋白是钩端螺旋体病检测的良好抗原。使用rLipL32的诊断试验是安全的,因为它不使用感染性细菌作为抗原,而且在印度尼西亚的每个诊断实验室都很容易进行。然而,rLipL32检测的现场验证还需要进一步的研究。
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