Elisa protocol for rapid screening of potential anti-tubercular drugs based on antigenic reactivity of mycobacterial ES-31 serine protease - a drug target supported by axenic culture of Mycobacterium tuberculosis H37 Ra strain in the presence of inhibitor
Mycobacterial ES-31 serine protease has been reported to be a drug target using protease and lipase inhibitors in axenic and macrophage cultures. Simple screening techniques are needed for rapid testing of anti-tubercular drugs. To demonstrate the usefulness of ELISA protocol based on antigenic reac...
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| Veröffentlicht in: | Indian journal of tuberculosis 2013-07, Vol.60 (3), p.138-141 |
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| creator | Hutke, Vinita Wankhade, Gauri Waghmare, Pranita J Harinath, B C |
| description | Mycobacterial ES-31 serine protease has been reported to be a drug target using protease and lipase inhibitors in axenic and macrophage cultures. Simple screening techniques are needed for rapid testing of anti-tubercular drugs.
To demonstrate the usefulness of ELISA protocol based on antigenic reactivity of mycobacterial serine protease by indirect ELISA for detecting anti-tubercular activity.
Indirect ELISA for assessment of antigenic reactivity of mycobacterial ES-31 serine protease was standardized using ES-31Ag and anti-DSS-goat-serum and assessed the inhibition of the antigenic reactivity by isoniazid, an anti-tubercular drug and serine protease inhibitor and orlistat, a lipase inhibitor.
Optimal antigenic reactivity of mycobacterial ES-31 serine protease was observed at 5 microg/well of ES-31 antigen and at 1:25 dilution of anti-DSS-goat-serum. Isoniazid showed 42% inhibition of ES-31 serine protease at 0.4 microg/well, while orlistat showed inhibition of 60% at 0.5 microg/well. Inhibition of Mtb H,37Ra bacilli is further confirmed in axenic culture. 35% and 29% inhibition by isoniazid at 0.4 microg/well and orlistat at 0.5 microg/well were observed respectively on bacterial growth.
Simple ELISA protocol based on assay of antigenic reactivity of mycobacterial ES-31 serine protease, a drug target, has been standardized for rapid screening of potential anti-tubercular drugs. |
| format | Article |
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To demonstrate the usefulness of ELISA protocol based on antigenic reactivity of mycobacterial serine protease by indirect ELISA for detecting anti-tubercular activity.
Indirect ELISA for assessment of antigenic reactivity of mycobacterial ES-31 serine protease was standardized using ES-31Ag and anti-DSS-goat-serum and assessed the inhibition of the antigenic reactivity by isoniazid, an anti-tubercular drug and serine protease inhibitor and orlistat, a lipase inhibitor.
Optimal antigenic reactivity of mycobacterial ES-31 serine protease was observed at 5 microg/well of ES-31 antigen and at 1:25 dilution of anti-DSS-goat-serum. Isoniazid showed 42% inhibition of ES-31 serine protease at 0.4 microg/well, while orlistat showed inhibition of 60% at 0.5 microg/well. Inhibition of Mtb H,37Ra bacilli is further confirmed in axenic culture. 35% and 29% inhibition by isoniazid at 0.4 microg/well and orlistat at 0.5 microg/well were observed respectively on bacterial growth.
Simple ELISA protocol based on assay of antigenic reactivity of mycobacterial ES-31 serine protease, a drug target, has been standardized for rapid screening of potential anti-tubercular drugs.</description><identifier>ISSN: 0019-5707</identifier><identifier>PMID: 24000490</identifier><language>eng</language><publisher>India</publisher><subject>Antitubercular Agents - pharmacokinetics ; Axenic Culture ; Bacterial Proteins - metabolism ; Drug Resistance, Microbial - physiology ; Enzyme-Linked Immunosorbent Assay - methods ; Humans ; Isoniazid - pharmacokinetics ; Lactones - pharmacokinetics ; Mycobacterium tuberculosis - drug effects ; Mycobacterium tuberculosis - metabolism ; Serine Proteases - metabolism ; Tuberculosis - drug therapy</subject><ispartof>Indian journal of tuberculosis, 2013-07, Vol.60 (3), p.138-141</ispartof><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/24000490$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Hutke, Vinita</creatorcontrib><creatorcontrib>Wankhade, Gauri</creatorcontrib><creatorcontrib>Waghmare, Pranita J</creatorcontrib><creatorcontrib>Harinath, B C</creatorcontrib><title>Elisa protocol for rapid screening of potential anti-tubercular drugs based on antigenic reactivity of mycobacterial ES-31 serine protease - a drug target supported by axenic culture of Mycobacterium tuberculosis H37 Ra strain in the presence of inhibitor</title><title>Indian journal of tuberculosis</title><addtitle>Indian J Tuberc</addtitle><description>Mycobacterial ES-31 serine protease has been reported to be a drug target using protease and lipase inhibitors in axenic and macrophage cultures. Simple screening techniques are needed for rapid testing of anti-tubercular drugs.
To demonstrate the usefulness of ELISA protocol based on antigenic reactivity of mycobacterial serine protease by indirect ELISA for detecting anti-tubercular activity.
Indirect ELISA for assessment of antigenic reactivity of mycobacterial ES-31 serine protease was standardized using ES-31Ag and anti-DSS-goat-serum and assessed the inhibition of the antigenic reactivity by isoniazid, an anti-tubercular drug and serine protease inhibitor and orlistat, a lipase inhibitor.
Optimal antigenic reactivity of mycobacterial ES-31 serine protease was observed at 5 microg/well of ES-31 antigen and at 1:25 dilution of anti-DSS-goat-serum. Isoniazid showed 42% inhibition of ES-31 serine protease at 0.4 microg/well, while orlistat showed inhibition of 60% at 0.5 microg/well. Inhibition of Mtb H,37Ra bacilli is further confirmed in axenic culture. 35% and 29% inhibition by isoniazid at 0.4 microg/well and orlistat at 0.5 microg/well were observed respectively on bacterial growth.
Simple ELISA protocol based on assay of antigenic reactivity of mycobacterial ES-31 serine protease, a drug target, has been standardized for rapid screening of potential anti-tubercular drugs.</description><subject>Antitubercular Agents - pharmacokinetics</subject><subject>Axenic Culture</subject><subject>Bacterial Proteins - metabolism</subject><subject>Drug Resistance, Microbial - physiology</subject><subject>Enzyme-Linked Immunosorbent Assay - methods</subject><subject>Humans</subject><subject>Isoniazid - pharmacokinetics</subject><subject>Lactones - pharmacokinetics</subject><subject>Mycobacterium tuberculosis - drug effects</subject><subject>Mycobacterium tuberculosis - metabolism</subject><subject>Serine Proteases - metabolism</subject><subject>Tuberculosis - drug therapy</subject><issn>0019-5707</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2013</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpFkU1PwzAMhnsA8TH4C8hHLpXSJmvXI0KDIQ0hAffJTdwR1CYlH4j9esgKE1IkK7bf503so-yMsaLJ5zWrT7Nz798Z44IJfpKdloIxJhp2ln0ve-0RRmeDlbaHzjpwOGoFXjoio80WbAejDWSCxh4whTzElpyMPTpQLm49tOhJgTVTeZtkEhyhDPpTh90eMOykbVOC3B6yfMl5AT5dDE3elPSQA044COi2FMDHcbQuJHC7A_yaqMk0REd75OM_Mg5weJL12sOK1_CM4INDbSCd8Lb3IU9GTlpt3nSrg3UX2XGHvafLvzjLXu-Wr7erfP10_3B7s87HecVygdgpWnSSCVyk2VVFU1WKN6LkVEqS9ZxLRk3ZoWBdmnqpFlhXVJVCFotOKT7Lrn-x6bMfkXzYDNpL6ns0ZKPfFIIz3tRcNKn16q81tgOpzej0gG63OeyM_wBK_Zhp</recordid><startdate>201307</startdate><enddate>201307</enddate><creator>Hutke, Vinita</creator><creator>Wankhade, Gauri</creator><creator>Waghmare, Pranita J</creator><creator>Harinath, B C</creator><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>7X8</scope></search><sort><creationdate>201307</creationdate><title>Elisa protocol for rapid screening of potential anti-tubercular drugs based on antigenic reactivity of mycobacterial ES-31 serine protease - a drug target supported by axenic culture of Mycobacterium tuberculosis H37 Ra strain in the presence of inhibitor</title><author>Hutke, Vinita ; Wankhade, Gauri ; Waghmare, Pranita J ; Harinath, B C</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-p560-4aafde8fc04a840061966d39423e2cec753c0e92fa40f0012d8a76e624c18fdd3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2013</creationdate><topic>Antitubercular Agents - pharmacokinetics</topic><topic>Axenic Culture</topic><topic>Bacterial Proteins - metabolism</topic><topic>Drug Resistance, Microbial - physiology</topic><topic>Enzyme-Linked Immunosorbent Assay - methods</topic><topic>Humans</topic><topic>Isoniazid - pharmacokinetics</topic><topic>Lactones - pharmacokinetics</topic><topic>Mycobacterium tuberculosis - drug effects</topic><topic>Mycobacterium tuberculosis - metabolism</topic><topic>Serine Proteases - metabolism</topic><topic>Tuberculosis - drug therapy</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Hutke, Vinita</creatorcontrib><creatorcontrib>Wankhade, Gauri</creatorcontrib><creatorcontrib>Waghmare, Pranita J</creatorcontrib><creatorcontrib>Harinath, B C</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>MEDLINE - Academic</collection><jtitle>Indian journal of tuberculosis</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Hutke, Vinita</au><au>Wankhade, Gauri</au><au>Waghmare, Pranita J</au><au>Harinath, B C</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Elisa protocol for rapid screening of potential anti-tubercular drugs based on antigenic reactivity of mycobacterial ES-31 serine protease - a drug target supported by axenic culture of Mycobacterium tuberculosis H37 Ra strain in the presence of inhibitor</atitle><jtitle>Indian journal of tuberculosis</jtitle><addtitle>Indian J Tuberc</addtitle><date>2013-07</date><risdate>2013</risdate><volume>60</volume><issue>3</issue><spage>138</spage><epage>141</epage><pages>138-141</pages><issn>0019-5707</issn><abstract>Mycobacterial ES-31 serine protease has been reported to be a drug target using protease and lipase inhibitors in axenic and macrophage cultures. Simple screening techniques are needed for rapid testing of anti-tubercular drugs.
To demonstrate the usefulness of ELISA protocol based on antigenic reactivity of mycobacterial serine protease by indirect ELISA for detecting anti-tubercular activity.
Indirect ELISA for assessment of antigenic reactivity of mycobacterial ES-31 serine protease was standardized using ES-31Ag and anti-DSS-goat-serum and assessed the inhibition of the antigenic reactivity by isoniazid, an anti-tubercular drug and serine protease inhibitor and orlistat, a lipase inhibitor.
Optimal antigenic reactivity of mycobacterial ES-31 serine protease was observed at 5 microg/well of ES-31 antigen and at 1:25 dilution of anti-DSS-goat-serum. Isoniazid showed 42% inhibition of ES-31 serine protease at 0.4 microg/well, while orlistat showed inhibition of 60% at 0.5 microg/well. Inhibition of Mtb H,37Ra bacilli is further confirmed in axenic culture. 35% and 29% inhibition by isoniazid at 0.4 microg/well and orlistat at 0.5 microg/well were observed respectively on bacterial growth.
Simple ELISA protocol based on assay of antigenic reactivity of mycobacterial ES-31 serine protease, a drug target, has been standardized for rapid screening of potential anti-tubercular drugs.</abstract><cop>India</cop><pmid>24000490</pmid><tpages>4</tpages></addata></record> |
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| ispartof | Indian journal of tuberculosis, 2013-07, Vol.60 (3), p.138-141 |
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| subjects | Antitubercular Agents - pharmacokinetics Axenic Culture Bacterial Proteins - metabolism Drug Resistance, Microbial - physiology Enzyme-Linked Immunosorbent Assay - methods Humans Isoniazid - pharmacokinetics Lactones - pharmacokinetics Mycobacterium tuberculosis - drug effects Mycobacterium tuberculosis - metabolism Serine Proteases - metabolism Tuberculosis - drug therapy |
| title | Elisa protocol for rapid screening of potential anti-tubercular drugs based on antigenic reactivity of mycobacterial ES-31 serine protease - a drug target supported by axenic culture of Mycobacterium tuberculosis H37 Ra strain in the presence of inhibitor |
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