Improved Antiserum for Sensitive Serologic Detection of Chickpea chlorotic dwarf virus

A Syrian chickpea isolate of Chickpea chlorotic dwarf virus (CpCDV; genus Mastrevirus, family Geminiviridae) was purified and yielded 0.6-0.8 mg of purified virus per kg of infected chickpea tissue. The purified preparations were injected into a rabbit and an antiserum of good quality was obtained a...

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Veröffentlicht in:Journal of phytopathology 2006-03, Vol.154 (3), p.129-133
Hauptverfasser: Kumari, S.G, Makkouk, K.M, Attar, N
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Attar, N
description A Syrian chickpea isolate of Chickpea chlorotic dwarf virus (CpCDV; genus Mastrevirus, family Geminiviridae) was purified and yielded 0.6-0.8 mg of purified virus per kg of infected chickpea tissue. The purified preparations were injected into a rabbit and an antiserum of good quality was obtained and used to evaluate different serological tests for the detection of CpCDV in infected chickpea leaf tissue and extracts. CpCDV was detected in sap dilutions of 1/640 by double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) and dot-blot ELISA, and in sap dilutions of 1/1280 by direct antigen-coating (DAC)-ELISA using CpCDV immunoglobulin G (IgG) at 0.5 [mu]g/ml. The antiserum was also able to detect the capsid protein of CpCDV by Western blot using raw antiserum at a dilution of 1/2000. The CpCDV raw antiserum (third bleeding) produced had a titre of 1/320 000 when determined by tissue-blot immunoassay (TBIA); whereas, coating ELISA plates with CpCDV IgG at a concentration of 0.004 [mu]g/ml was enough to detect the virus by DAS-ELISA in a sap dilution of 1/20 using an enzyme conjugate at a dilution of 1/2000.
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The CpCDV raw antiserum (third bleeding) produced had a titre of 1/320 000 when determined by tissue-blot immunoassay (TBIA); whereas, coating ELISA plates with CpCDV IgG at a concentration of 0.004 [mu]g/ml was enough to detect the virus by DAS-ELISA in a sap dilution of 1/20 using an enzyme conjugate at a dilution of 1/2000.</description><identifier>ISSN: 0931-1785</identifier><identifier>EISSN: 1439-0434</identifier><identifier>DOI: 10.1111/j.1439-0434.2006.01068.x</identifier><identifier>CODEN: JPHYEB</identifier><language>eng</language><publisher>Berlin, Germany: Berlin, Germany : Blackwell Verlag GmbH</publisher><subject>antiserum ; Biological and medical sciences ; Chickpea chlorotic dwarf virus ; chickpeas ; Cicer arietinum ; coat proteins ; diagnosis ; direct antigen-coating enzyme-linked immunosorbent assay ; disease detection ; disease diagnosis ; dot-blot ; double-antibody sandwich enzyme-linked immunosorbent assay ; enzyme-linked immunosorbent assay ; Fundamental and applied biological sciences. Psychology ; Geminivirus ; immunologic techniques ; leaves ; Mastrevirus ; new methods ; Phytopathology. Animal pests. 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The purified preparations were injected into a rabbit and an antiserum of good quality was obtained and used to evaluate different serological tests for the detection of CpCDV in infected chickpea leaf tissue and extracts. CpCDV was detected in sap dilutions of 1/640 by double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) and dot-blot ELISA, and in sap dilutions of 1/1280 by direct antigen-coating (DAC)-ELISA using CpCDV immunoglobulin G (IgG) at 0.5 [mu]g/ml. The antiserum was also able to detect the capsid protein of CpCDV by Western blot using raw antiserum at a dilution of 1/2000. The CpCDV raw antiserum (third bleeding) produced had a titre of 1/320 000 when determined by tissue-blot immunoassay (TBIA); whereas, coating ELISA plates with CpCDV IgG at a concentration of 0.004 [mu]g/ml was enough to detect the virus by DAS-ELISA in a sap dilution of 1/20 using an enzyme conjugate at a dilution of 1/2000.</description><subject>antiserum</subject><subject>Biological and medical sciences</subject><subject>Chickpea chlorotic dwarf virus</subject><subject>chickpeas</subject><subject>Cicer arietinum</subject><subject>coat proteins</subject><subject>diagnosis</subject><subject>direct antigen-coating enzyme-linked immunosorbent assay</subject><subject>disease detection</subject><subject>disease diagnosis</subject><subject>dot-blot</subject><subject>double-antibody sandwich enzyme-linked immunosorbent assay</subject><subject>enzyme-linked immunosorbent assay</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Geminivirus</subject><subject>immunologic techniques</subject><subject>leaves</subject><subject>Mastrevirus</subject><subject>new methods</subject><subject>Phytopathology. Animal pests. Plant and forest protection</subject><subject>plant extracts</subject><subject>plant viruses</subject><subject>Plant viruses and viroids</subject><subject>sap</subject><subject>serodiagnosis</subject><subject>tissue-blot immunoassay</subject><subject>Western blot</subject><issn>0931-1785</issn><issn>1439-0434</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2006</creationdate><recordtype>article</recordtype><recordid>eNqNkE9v1DAQxS0EEkvhM-AL3BLGdhzbBw7VQv9RFaRSOFquY7feZuOtnd1uvz0OqcqVucxI83tv7IcQJlCTUp9WNWmYqqBhTU0B2hoItLLev0CL58VLtADFSEWE5K_Rm5xXABQYwAL9Ol1vUty5Dh8OY8gubdfYx4Qv3ZDDGHauTCn28SZY_MWNzo4hDjh6vLwN9m7jDLa3fUxxLPvuwSSPdyFt81v0yps-u3dP_QBdHX39uTypzr8fny4PzyvbsFZWXlBLjSINaToruLz2SvAOjLyW1DhnGNjWMdUa01ghlBQdKMmNY0IqaqFjB-jj7Fs-cb91edTrkK3rezO4uM2aCMI5IbyAcgZtijkn5_UmhbVJj5qAnoLUKz3lpae89BSk_huk3hfph6cbJlvT-2QGG_I_veDAmKCF-zxzD6F3j__tr89-nExT0VezPuTR7Z_1Jt3pVjDB9e-LY33WUvbt6ILrZeHfz7w3UZubVN50dUmBsGLMJSjB_gCLAp7T</recordid><startdate>200603</startdate><enddate>200603</enddate><creator>Kumari, S.G</creator><creator>Makkouk, K.M</creator><creator>Attar, N</creator><general>Berlin, Germany : Blackwell Verlag GmbH</general><general>Blackwell Verlag GmbH</general><general>Blackwell</general><scope>FBQ</scope><scope>BSCLL</scope><scope>IQODW</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7U9</scope><scope>H94</scope></search><sort><creationdate>200603</creationdate><title>Improved Antiserum for Sensitive Serologic Detection of Chickpea chlorotic dwarf virus</title><author>Kumari, S.G ; Makkouk, K.M ; Attar, N</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4368-f72c2a91414dc758bf975d0a8b82aeea30c6e396aa4c77987d0985ae37892c0d3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2006</creationdate><topic>antiserum</topic><topic>Biological and medical sciences</topic><topic>Chickpea chlorotic dwarf virus</topic><topic>chickpeas</topic><topic>Cicer arietinum</topic><topic>coat proteins</topic><topic>diagnosis</topic><topic>direct antigen-coating enzyme-linked immunosorbent assay</topic><topic>disease detection</topic><topic>disease diagnosis</topic><topic>dot-blot</topic><topic>double-antibody sandwich enzyme-linked immunosorbent assay</topic><topic>enzyme-linked immunosorbent assay</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Geminivirus</topic><topic>immunologic techniques</topic><topic>leaves</topic><topic>Mastrevirus</topic><topic>new methods</topic><topic>Phytopathology. Animal pests. Plant and forest protection</topic><topic>plant extracts</topic><topic>plant viruses</topic><topic>Plant viruses and viroids</topic><topic>sap</topic><topic>serodiagnosis</topic><topic>tissue-blot immunoassay</topic><topic>Western blot</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kumari, S.G</creatorcontrib><creatorcontrib>Makkouk, K.M</creatorcontrib><creatorcontrib>Attar, N</creatorcontrib><collection>AGRIS</collection><collection>Istex</collection><collection>Pascal-Francis</collection><collection>CrossRef</collection><collection>Virology and AIDS Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><jtitle>Journal of phytopathology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kumari, S.G</au><au>Makkouk, K.M</au><au>Attar, N</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Improved Antiserum for Sensitive Serologic Detection of Chickpea chlorotic dwarf virus</atitle><jtitle>Journal of phytopathology</jtitle><date>2006-03</date><risdate>2006</risdate><volume>154</volume><issue>3</issue><spage>129</spage><epage>133</epage><pages>129-133</pages><issn>0931-1785</issn><eissn>1439-0434</eissn><coden>JPHYEB</coden><abstract>A Syrian chickpea isolate of Chickpea chlorotic dwarf virus (CpCDV; genus Mastrevirus, family Geminiviridae) was purified and yielded 0.6-0.8 mg of purified virus per kg of infected chickpea tissue. The purified preparations were injected into a rabbit and an antiserum of good quality was obtained and used to evaluate different serological tests for the detection of CpCDV in infected chickpea leaf tissue and extracts. CpCDV was detected in sap dilutions of 1/640 by double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) and dot-blot ELISA, and in sap dilutions of 1/1280 by direct antigen-coating (DAC)-ELISA using CpCDV immunoglobulin G (IgG) at 0.5 [mu]g/ml. The antiserum was also able to detect the capsid protein of CpCDV by Western blot using raw antiserum at a dilution of 1/2000. The CpCDV raw antiserum (third bleeding) produced had a titre of 1/320 000 when determined by tissue-blot immunoassay (TBIA); whereas, coating ELISA plates with CpCDV IgG at a concentration of 0.004 [mu]g/ml was enough to detect the virus by DAS-ELISA in a sap dilution of 1/20 using an enzyme conjugate at a dilution of 1/2000.</abstract><cop>Berlin, Germany</cop><pub>Berlin, Germany : Blackwell Verlag GmbH</pub><doi>10.1111/j.1439-0434.2006.01068.x</doi><tpages>5</tpages></addata></record>
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subjects antiserum
Biological and medical sciences
Chickpea chlorotic dwarf virus
chickpeas
Cicer arietinum
coat proteins
diagnosis
direct antigen-coating enzyme-linked immunosorbent assay
disease detection
disease diagnosis
dot-blot
double-antibody sandwich enzyme-linked immunosorbent assay
enzyme-linked immunosorbent assay
Fundamental and applied biological sciences. Psychology
Geminivirus
immunologic techniques
leaves
Mastrevirus
new methods
Phytopathology. Animal pests. Plant and forest protection
plant extracts
plant viruses
Plant viruses and viroids
sap
serodiagnosis
tissue-blot immunoassay
Western blot
title Improved Antiserum for Sensitive Serologic Detection of Chickpea chlorotic dwarf virus
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