{"title":"Recombinant LipL32 Protein for Leptospirosis Detection in Indonesia","authors":"Sumarningsih, Simson Tarigan, Susanti, Kusmiyati","doi":"10.1016/j.proche.2016.01.004","DOIUrl":null,"url":null,"abstract":"<div><p>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 <em>E. coli</em> 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.</p></div>","PeriodicalId":20431,"journal":{"name":"Procedia Chemistry","volume":"18 ","pages":"Pages 18-25"},"PeriodicalIF":0.0000,"publicationDate":"2016-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.proche.2016.01.004","citationCount":"8","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Procedia Chemistry","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S187661961600005X","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
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.