New Sandwich-Type Enzyme-Linked Immunosorbent Assay for Human MxA Protein in a Whole Blood Using Monoclonal Antibodies Against GTP-Binding Domain for Recognition of Viral Infection
Objectives To develop a clinically significant and practical enzyme‐linked immunosorbent assay (ELISA) for the detection of MxA protein in human whole blood, a biological marker of viral infection. Design and Methods A sandwich ELISA suitable for the measurement of human MxA protein in whole blood w...
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| Veröffentlicht in: | Journal of clinical laboratory analysis 2012-05, Vol.26 (3), p.174-183 |
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| creator | Kawamura, Mizuho Kusano, Akira Furuya, Akiko Hanai, Nobuo Tanigaki, Hideki Tomita, Akihito Horiguchi, Akira Nagata, Kyosuke Itazawa, Toshiko Adachi, Yuichi Okabe, Yoshie Miyawaki, Toshio Kohno, Hiroaki |
| description | Objectives
To develop a clinically significant and practical enzyme‐linked immunosorbent assay (ELISA) for the detection of MxA protein in human whole blood, a biological marker of viral infection.
Design and Methods
A sandwich ELISA suitable for the measurement of human MxA protein in whole blood was developed using mouse monoclonal antibodies (mAbs) raised against the GTP‐binding domain of human MxA protein. Prior to the assay, the whole blood sample was treated with special buffer to extract the MxA protein, improve its stability, and avoid interference from hemoglobin.
Results
This ELISA meets all the requirements for use in routine clinical assays, especially in terms of sensitivity (detection limit: 1.3 ng/ml whole blood), accuracy (recovery: 93.0–100.0%), and rapidity ( |
| doi_str_mv | 10.1002/jcla.21507 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_pubme</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_6807620</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>1016670274</sourcerecordid><originalsourceid>FETCH-LOGICAL-c5857-19ffc31d7f0c7b34b443d75a7c809ca4f1ba148d77fda6d12b5dfee68c0cd7723</originalsourceid><addsrcrecordid>eNqFkt9u0zAUhyMEYmVwwwMgS9wgpAzb-WP3Bikro-voxjQ6tjvLsZ3WXWIXO6Erz8UD4qxbBVyAZMnS8Xc-HR_9ouglggcIQvxuKWp-gFEGyaNogOCQxpji7HE0gJSSmEKU7EXPvF9CCOkQ5U-jPYzzgCTJIPp5ptbgCzdyrcUinm1WChyZH5tGxVNtbpQEk6bpjPXWlcq0oPCeb0BlHTjuGm7A6W0Bzp1tlTYgHA6uFrZW4LC2VoJLr80cnFpjRW0Nr0FhWl1aqZUHxZxr41swnp3Hh9rInvxgm1C8s18oYedGt9oaYCvwVbvQPjGVEn3pefSk4rVXL-7v_ejy49FsdBxPP48no2Iai4xmJEbDqhIJkqSCgpRJWqZpIknGiaBwKHhaoZKjlEpCKslziXCZyUqpnAooQhEn-9H7rXfVlY2SImwgzMFWTjfcbZjlmv35YvSCze13llNIcgyD4M29wNlvnfIta7QXqq65UbbzDKUpxThN8-H_UYjynEBM0oC-_gtd2s6FBffCJMdphnMSqLdbSjjrvVPVbm4EWZ8b1ueG3eUmwK9-_-kOfQhKANAWWOtabf6hYiejafEgjbc92rfqdtfD3Q0L85GMXZ2N2fU1-nRxgjKGkl8Cx99B</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1436245267</pqid></control><display><type>article</type><title>New Sandwich-Type Enzyme-Linked Immunosorbent Assay for Human MxA Protein in a Whole Blood Using Monoclonal Antibodies Against GTP-Binding Domain for Recognition of Viral Infection</title><source>MEDLINE</source><source>Wiley Journals</source><source>EZB-FREE-00999 freely available EZB journals</source><source>PubMed Central</source><creator>Kawamura, Mizuho ; Kusano, Akira ; Furuya, Akiko ; Hanai, Nobuo ; Tanigaki, Hideki ; Tomita, Akihito ; Horiguchi, Akira ; Nagata, Kyosuke ; Itazawa, Toshiko ; Adachi, Yuichi ; Okabe, Yoshie ; Miyawaki, Toshio ; Kohno, Hiroaki</creator><creatorcontrib>Kawamura, Mizuho ; Kusano, Akira ; Furuya, Akiko ; Hanai, Nobuo ; Tanigaki, Hideki ; Tomita, Akihito ; Horiguchi, Akira ; Nagata, Kyosuke ; Itazawa, Toshiko ; Adachi, Yuichi ; Okabe, Yoshie ; Miyawaki, Toshio ; Kohno, Hiroaki</creatorcontrib><description>Objectives
To develop a clinically significant and practical enzyme‐linked immunosorbent assay (ELISA) for the detection of MxA protein in human whole blood, a biological marker of viral infection.
Design and Methods
A sandwich ELISA suitable for the measurement of human MxA protein in whole blood was developed using mouse monoclonal antibodies (mAbs) raised against the GTP‐binding domain of human MxA protein. Prior to the assay, the whole blood sample was treated with special buffer to extract the MxA protein, improve its stability, and avoid interference from hemoglobin.
Results
This ELISA meets all the requirements for use in routine clinical assays, especially in terms of sensitivity (detection limit: 1.3 ng/ml whole blood), accuracy (recovery: 93.0–100.0%), and rapidity (<1.5 h). The present ELISA had a sensitivity of100% and a specificity of 100% for viral infection when compared to samples from healthy control and 87.1% and 90.9% when compared to samples from the bacterial infection group.
Conclusion
We have developed a new ELISA for measuring MxA protein in human whole blood using mAbs specific for the GTP‐binding domain of MxA. This ELISA has analytical performance enough for routine clinical assay and can be used in detecting the possibility of viral infection. J. Clin. Lab. Anal. 26:174‐183, 2012. © 2012 Wiley Periodicals, Inc.</description><identifier>ISSN: 0887-8013</identifier><identifier>EISSN: 1098-2825</identifier><identifier>DOI: 10.1002/jcla.21507</identifier><identifier>PMID: 22628233</identifier><language>eng</language><publisher>United States: Blackwell Publishing Ltd</publisher><subject>Animals ; Antibodies, Monoclonal - chemistry ; Antibodies, Monoclonal - immunology ; bacterial infection ; Bacterial Infections - blood ; Bacterial Infections - diagnosis ; Binding Sites ; Biomarkers - blood ; Case-Control Studies ; Child ; Child, Preschool ; Enzyme-Linked Immunosorbent Assay - methods ; Enzymes ; Female ; GTP-Binding Proteins - blood ; GTP-Binding Proteins - immunology ; high sensitivity ; high stability ; Humans ; Immunoassay ; Infant ; interferon α ; interferon α, high sensitivity ; Male ; Mice ; Monoclonal antibodies ; Myxovirus Resistance Proteins ; Protein Stability ; Proteins ; rapid assay ; Sensitivity and Specificity ; Viral infections ; Virus Diseases - blood ; Virus Diseases - diagnosis</subject><ispartof>Journal of clinical laboratory analysis, 2012-05, Vol.26 (3), p.174-183</ispartof><rights>2012 Wiley Periodicals, Inc.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c5857-19ffc31d7f0c7b34b443d75a7c809ca4f1ba148d77fda6d12b5dfee68c0cd7723</citedby><cites>FETCH-LOGICAL-c5857-19ffc31d7f0c7b34b443d75a7c809ca4f1ba148d77fda6d12b5dfee68c0cd7723</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6807620/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC6807620/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,727,780,784,885,1417,27924,27925,45574,45575,53791,53793</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/22628233$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Kawamura, Mizuho</creatorcontrib><creatorcontrib>Kusano, Akira</creatorcontrib><creatorcontrib>Furuya, Akiko</creatorcontrib><creatorcontrib>Hanai, Nobuo</creatorcontrib><creatorcontrib>Tanigaki, Hideki</creatorcontrib><creatorcontrib>Tomita, Akihito</creatorcontrib><creatorcontrib>Horiguchi, Akira</creatorcontrib><creatorcontrib>Nagata, Kyosuke</creatorcontrib><creatorcontrib>Itazawa, Toshiko</creatorcontrib><creatorcontrib>Adachi, Yuichi</creatorcontrib><creatorcontrib>Okabe, Yoshie</creatorcontrib><creatorcontrib>Miyawaki, Toshio</creatorcontrib><creatorcontrib>Kohno, Hiroaki</creatorcontrib><title>New Sandwich-Type Enzyme-Linked Immunosorbent Assay for Human MxA Protein in a Whole Blood Using Monoclonal Antibodies Against GTP-Binding Domain for Recognition of Viral Infection</title><title>Journal of clinical laboratory analysis</title><addtitle>J. Clin. Lab. Anal</addtitle><description>Objectives
To develop a clinically significant and practical enzyme‐linked immunosorbent assay (ELISA) for the detection of MxA protein in human whole blood, a biological marker of viral infection.
Design and Methods
A sandwich ELISA suitable for the measurement of human MxA protein in whole blood was developed using mouse monoclonal antibodies (mAbs) raised against the GTP‐binding domain of human MxA protein. Prior to the assay, the whole blood sample was treated with special buffer to extract the MxA protein, improve its stability, and avoid interference from hemoglobin.
Results
This ELISA meets all the requirements for use in routine clinical assays, especially in terms of sensitivity (detection limit: 1.3 ng/ml whole blood), accuracy (recovery: 93.0–100.0%), and rapidity (<1.5 h). The present ELISA had a sensitivity of100% and a specificity of 100% for viral infection when compared to samples from healthy control and 87.1% and 90.9% when compared to samples from the bacterial infection group.
Conclusion
We have developed a new ELISA for measuring MxA protein in human whole blood using mAbs specific for the GTP‐binding domain of MxA. This ELISA has analytical performance enough for routine clinical assay and can be used in detecting the possibility of viral infection. J. Clin. Lab. Anal. 26:174‐183, 2012. © 2012 Wiley Periodicals, Inc.</description><subject>Animals</subject><subject>Antibodies, Monoclonal - chemistry</subject><subject>Antibodies, Monoclonal - immunology</subject><subject>bacterial infection</subject><subject>Bacterial Infections - blood</subject><subject>Bacterial Infections - diagnosis</subject><subject>Binding Sites</subject><subject>Biomarkers - blood</subject><subject>Case-Control Studies</subject><subject>Child</subject><subject>Child, Preschool</subject><subject>Enzyme-Linked Immunosorbent Assay - methods</subject><subject>Enzymes</subject><subject>Female</subject><subject>GTP-Binding Proteins - blood</subject><subject>GTP-Binding Proteins - immunology</subject><subject>high sensitivity</subject><subject>high stability</subject><subject>Humans</subject><subject>Immunoassay</subject><subject>Infant</subject><subject>interferon α</subject><subject>interferon α, high sensitivity</subject><subject>Male</subject><subject>Mice</subject><subject>Monoclonal antibodies</subject><subject>Myxovirus Resistance Proteins</subject><subject>Protein Stability</subject><subject>Proteins</subject><subject>rapid assay</subject><subject>Sensitivity and Specificity</subject><subject>Viral infections</subject><subject>Virus Diseases - blood</subject><subject>Virus Diseases - diagnosis</subject><issn>0887-8013</issn><issn>1098-2825</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2012</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkt9u0zAUhyMEYmVwwwMgS9wgpAzb-WP3Bikro-voxjQ6tjvLsZ3WXWIXO6Erz8UD4qxbBVyAZMnS8Xc-HR_9ouglggcIQvxuKWp-gFEGyaNogOCQxpji7HE0gJSSmEKU7EXPvF9CCOkQ5U-jPYzzgCTJIPp5ptbgCzdyrcUinm1WChyZH5tGxVNtbpQEk6bpjPXWlcq0oPCeb0BlHTjuGm7A6W0Bzp1tlTYgHA6uFrZW4LC2VoJLr80cnFpjRW0Nr0FhWl1aqZUHxZxr41swnp3Hh9rInvxgm1C8s18oYedGt9oaYCvwVbvQPjGVEn3pefSk4rVXL-7v_ejy49FsdBxPP48no2Iai4xmJEbDqhIJkqSCgpRJWqZpIknGiaBwKHhaoZKjlEpCKslziXCZyUqpnAooQhEn-9H7rXfVlY2SImwgzMFWTjfcbZjlmv35YvSCze13llNIcgyD4M29wNlvnfIta7QXqq65UbbzDKUpxThN8-H_UYjynEBM0oC-_gtd2s6FBffCJMdphnMSqLdbSjjrvVPVbm4EWZ8b1ueG3eUmwK9-_-kOfQhKANAWWOtabf6hYiejafEgjbc92rfqdtfD3Q0L85GMXZ2N2fU1-nRxgjKGkl8Cx99B</recordid><startdate>201205</startdate><enddate>201205</enddate><creator>Kawamura, Mizuho</creator><creator>Kusano, Akira</creator><creator>Furuya, Akiko</creator><creator>Hanai, Nobuo</creator><creator>Tanigaki, Hideki</creator><creator>Tomita, Akihito</creator><creator>Horiguchi, Akira</creator><creator>Nagata, Kyosuke</creator><creator>Itazawa, Toshiko</creator><creator>Adachi, Yuichi</creator><creator>Okabe, Yoshie</creator><creator>Miyawaki, Toshio</creator><creator>Kohno, Hiroaki</creator><general>Blackwell Publishing Ltd</general><general>John Wiley & Sons, Inc</general><general>John Wiley and Sons Inc</general><scope>BSCLL</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QP</scope><scope>7T5</scope><scope>7U9</scope><scope>H94</scope><scope>K9.</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>201205</creationdate><title>New Sandwich-Type Enzyme-Linked Immunosorbent Assay for Human MxA Protein in a Whole Blood Using Monoclonal Antibodies Against GTP-Binding Domain for Recognition of Viral Infection</title><author>Kawamura, Mizuho ; Kusano, Akira ; Furuya, Akiko ; Hanai, Nobuo ; Tanigaki, Hideki ; Tomita, Akihito ; Horiguchi, Akira ; Nagata, Kyosuke ; Itazawa, Toshiko ; Adachi, Yuichi ; Okabe, Yoshie ; Miyawaki, Toshio ; Kohno, Hiroaki</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c5857-19ffc31d7f0c7b34b443d75a7c809ca4f1ba148d77fda6d12b5dfee68c0cd7723</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2012</creationdate><topic>Animals</topic><topic>Antibodies, Monoclonal - chemistry</topic><topic>Antibodies, Monoclonal - immunology</topic><topic>bacterial infection</topic><topic>Bacterial Infections - blood</topic><topic>Bacterial Infections - diagnosis</topic><topic>Binding Sites</topic><topic>Biomarkers - blood</topic><topic>Case-Control Studies</topic><topic>Child</topic><topic>Child, Preschool</topic><topic>Enzyme-Linked Immunosorbent Assay - methods</topic><topic>Enzymes</topic><topic>Female</topic><topic>GTP-Binding Proteins - blood</topic><topic>GTP-Binding Proteins - immunology</topic><topic>high sensitivity</topic><topic>high stability</topic><topic>Humans</topic><topic>Immunoassay</topic><topic>Infant</topic><topic>interferon α</topic><topic>interferon α, high sensitivity</topic><topic>Male</topic><topic>Mice</topic><topic>Monoclonal antibodies</topic><topic>Myxovirus Resistance Proteins</topic><topic>Protein Stability</topic><topic>Proteins</topic><topic>rapid assay</topic><topic>Sensitivity and Specificity</topic><topic>Viral infections</topic><topic>Virus Diseases - blood</topic><topic>Virus Diseases - diagnosis</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kawamura, Mizuho</creatorcontrib><creatorcontrib>Kusano, Akira</creatorcontrib><creatorcontrib>Furuya, Akiko</creatorcontrib><creatorcontrib>Hanai, Nobuo</creatorcontrib><creatorcontrib>Tanigaki, Hideki</creatorcontrib><creatorcontrib>Tomita, Akihito</creatorcontrib><creatorcontrib>Horiguchi, Akira</creatorcontrib><creatorcontrib>Nagata, Kyosuke</creatorcontrib><creatorcontrib>Itazawa, Toshiko</creatorcontrib><creatorcontrib>Adachi, Yuichi</creatorcontrib><creatorcontrib>Okabe, Yoshie</creatorcontrib><creatorcontrib>Miyawaki, Toshio</creatorcontrib><creatorcontrib>Kohno, Hiroaki</creatorcontrib><collection>Istex</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Immunology Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Journal of clinical laboratory analysis</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kawamura, Mizuho</au><au>Kusano, Akira</au><au>Furuya, Akiko</au><au>Hanai, Nobuo</au><au>Tanigaki, Hideki</au><au>Tomita, Akihito</au><au>Horiguchi, Akira</au><au>Nagata, Kyosuke</au><au>Itazawa, Toshiko</au><au>Adachi, Yuichi</au><au>Okabe, Yoshie</au><au>Miyawaki, Toshio</au><au>Kohno, Hiroaki</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>New Sandwich-Type Enzyme-Linked Immunosorbent Assay for Human MxA Protein in a Whole Blood Using Monoclonal Antibodies Against GTP-Binding Domain for Recognition of Viral Infection</atitle><jtitle>Journal of clinical laboratory analysis</jtitle><addtitle>J. Clin. Lab. Anal</addtitle><date>2012-05</date><risdate>2012</risdate><volume>26</volume><issue>3</issue><spage>174</spage><epage>183</epage><pages>174-183</pages><issn>0887-8013</issn><eissn>1098-2825</eissn><abstract>Objectives
To develop a clinically significant and practical enzyme‐linked immunosorbent assay (ELISA) for the detection of MxA protein in human whole blood, a biological marker of viral infection.
Design and Methods
A sandwich ELISA suitable for the measurement of human MxA protein in whole blood was developed using mouse monoclonal antibodies (mAbs) raised against the GTP‐binding domain of human MxA protein. Prior to the assay, the whole blood sample was treated with special buffer to extract the MxA protein, improve its stability, and avoid interference from hemoglobin.
Results
This ELISA meets all the requirements for use in routine clinical assays, especially in terms of sensitivity (detection limit: 1.3 ng/ml whole blood), accuracy (recovery: 93.0–100.0%), and rapidity (<1.5 h). The present ELISA had a sensitivity of100% and a specificity of 100% for viral infection when compared to samples from healthy control and 87.1% and 90.9% when compared to samples from the bacterial infection group.
Conclusion
We have developed a new ELISA for measuring MxA protein in human whole blood using mAbs specific for the GTP‐binding domain of MxA. This ELISA has analytical performance enough for routine clinical assay and can be used in detecting the possibility of viral infection. J. Clin. Lab. Anal. 26:174‐183, 2012. © 2012 Wiley Periodicals, Inc.</abstract><cop>United States</cop><pub>Blackwell Publishing Ltd</pub><pmid>22628233</pmid><doi>10.1002/jcla.21507</doi><tpages>10</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Journal of clinical laboratory analysis, 2012-05, Vol.26 (3), p.174-183 |
| issn | 0887-8013 1098-2825 |
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| source | MEDLINE; Wiley Journals; EZB-FREE-00999 freely available EZB journals; PubMed Central |
| subjects | Animals Antibodies, Monoclonal - chemistry Antibodies, Monoclonal - immunology bacterial infection Bacterial Infections - blood Bacterial Infections - diagnosis Binding Sites Biomarkers - blood Case-Control Studies Child Child, Preschool Enzyme-Linked Immunosorbent Assay - methods Enzymes Female GTP-Binding Proteins - blood GTP-Binding Proteins - immunology high sensitivity high stability Humans Immunoassay Infant interferon α interferon α, high sensitivity Male Mice Monoclonal antibodies Myxovirus Resistance Proteins Protein Stability Proteins rapid assay Sensitivity and Specificity Viral infections Virus Diseases - blood Virus Diseases - diagnosis |
| title | New Sandwich-Type Enzyme-Linked Immunosorbent Assay for Human MxA Protein in a Whole Blood Using Monoclonal Antibodies Against GTP-Binding Domain for Recognition of Viral Infection |
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