The development of an automated in situ assay for the detection of human cytomegalovirus in peripheral blood leukocytes
Human cytomegalovirus (HCMV) infections are common in immunosuppressed patients, especially transplant recipients and patients with AIDS. The utility of an automated in situ hybridization (ISH) assay for the rapid detection of HCMV immediate early mRNA was evaluated using cytospin (Shandon Lipshaw,...
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| Veröffentlicht in: | Journal of virological methods 1999-08, Vol.81 (1), p.31-37 |
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| creator | Baunoch, David A. Lobell, Allegra Kane, Dorothy Fredericks, Judith Thompson, Kenneth D. |
| description | Human cytomegalovirus (HCMV) infections are common in immunosuppressed patients, especially transplant recipients and patients with AIDS. The utility of an automated in situ hybridization (ISH) assay for the rapid detection of HCMV immediate early mRNA was evaluated using cytospin (Shandon Lipshaw, Inc., Pittsburgh, PA) prepared leukocytes from peripheral blood samples. In this study, the detection of HCMV immediate early protein by immunofluorescent antibody staining of the standard shell vial assay was compared to the detection of HCMV immediate early mRNA in peripheral blood leukocytes using the automated ISH system. Of 135 specimens tested, eight specimens were positive using HCMV ISH compared to seven positive specimens using shell vial assay. Overall, HCMV ISH demonstrated 100% sensitivity and 99% specificity. Since the HCMV ISH assay requires minimal labor, and can be completed in less than 5 h, this method should be evaluated as a potential replacement for shell vial assay for the diagnosis of HCMV infection. |
| doi_str_mv | 10.1016/S0166-0934(99)00062-2 |
| format | Article |
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The utility of an automated in situ hybridization (ISH) assay for the rapid detection of HCMV immediate early mRNA was evaluated using cytospin (Shandon Lipshaw, Inc., Pittsburgh, PA) prepared leukocytes from peripheral blood samples. In this study, the detection of HCMV immediate early protein by immunofluorescent antibody staining of the standard shell vial assay was compared to the detection of HCMV immediate early mRNA in peripheral blood leukocytes using the automated ISH system. Of 135 specimens tested, eight specimens were positive using HCMV ISH compared to seven positive specimens using shell vial assay. Overall, HCMV ISH demonstrated 100% sensitivity and 99% specificity. Since the HCMV ISH assay requires minimal labor, and can be completed in less than 5 h, this method should be evaluated as a potential replacement for shell vial assay for the diagnosis of HCMV infection.</description><identifier>ISSN: 0166-0934</identifier><identifier>EISSN: 1879-0984</identifier><identifier>DOI: 10.1016/S0166-0934(99)00062-2</identifier><identifier>PMID: 10488758</identifier><identifier>CODEN: JVMEDH</identifier><language>eng</language><publisher>London: Elsevier B.V</publisher><subject>Biological and medical sciences ; Cytomegalovirus ; Cytomegalovirus - genetics ; Cytomegalovirus - isolation & purification ; Fibroblasts - chemistry ; Fibroblasts - virology ; Fundamental and applied biological sciences. Psychology ; Human cytomegalovirus ; Human viral diseases ; Humans ; Immediate early mRNA ; Immediate-Early Proteins - genetics ; In situ hybridization ; In Situ Hybridization - instrumentation ; In Situ Hybridization - methods ; Infectious diseases ; Leukocytes ; Leukocytes - chemistry ; Leukocytes - virology ; Medical sciences ; Microbiology ; RNA, Messenger - genetics ; RNA, Viral - genetics ; Techniques used in virology ; Viral diseases ; Viral diseases of the lymphoid tissue and the blood. 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The utility of an automated in situ hybridization (ISH) assay for the rapid detection of HCMV immediate early mRNA was evaluated using cytospin (Shandon Lipshaw, Inc., Pittsburgh, PA) prepared leukocytes from peripheral blood samples. In this study, the detection of HCMV immediate early protein by immunofluorescent antibody staining of the standard shell vial assay was compared to the detection of HCMV immediate early mRNA in peripheral blood leukocytes using the automated ISH system. Of 135 specimens tested, eight specimens were positive using HCMV ISH compared to seven positive specimens using shell vial assay. Overall, HCMV ISH demonstrated 100% sensitivity and 99% specificity. Since the HCMV ISH assay requires minimal labor, and can be completed in less than 5 h, this method should be evaluated as a potential replacement for shell vial assay for the diagnosis of HCMV infection.</description><subject>Biological and medical sciences</subject><subject>Cytomegalovirus</subject><subject>Cytomegalovirus - genetics</subject><subject>Cytomegalovirus - isolation & purification</subject><subject>Fibroblasts - chemistry</subject><subject>Fibroblasts - virology</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Human cytomegalovirus</subject><subject>Human viral diseases</subject><subject>Humans</subject><subject>Immediate early mRNA</subject><subject>Immediate-Early Proteins - genetics</subject><subject>In situ hybridization</subject><subject>In Situ Hybridization - instrumentation</subject><subject>In Situ Hybridization - methods</subject><subject>Infectious diseases</subject><subject>Leukocytes</subject><subject>Leukocytes - chemistry</subject><subject>Leukocytes - virology</subject><subject>Medical sciences</subject><subject>Microbiology</subject><subject>RNA, Messenger - genetics</subject><subject>RNA, Viral - genetics</subject><subject>Techniques used in virology</subject><subject>Viral diseases</subject><subject>Viral diseases of the lymphoid tissue and the blood. 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Psychology</topic><topic>Human cytomegalovirus</topic><topic>Human viral diseases</topic><topic>Humans</topic><topic>Immediate early mRNA</topic><topic>Immediate-Early Proteins - genetics</topic><topic>In situ hybridization</topic><topic>In Situ Hybridization - instrumentation</topic><topic>In Situ Hybridization - methods</topic><topic>Infectious diseases</topic><topic>Leukocytes</topic><topic>Leukocytes - chemistry</topic><topic>Leukocytes - virology</topic><topic>Medical sciences</topic><topic>Microbiology</topic><topic>RNA, Messenger - genetics</topic><topic>RNA, Viral - genetics</topic><topic>Techniques used in virology</topic><topic>Viral diseases</topic><topic>Viral diseases of the lymphoid tissue and the blood. Aids</topic><topic>Virology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Baunoch, David A.</creatorcontrib><creatorcontrib>Lobell, Allegra</creatorcontrib><creatorcontrib>Kane, Dorothy</creatorcontrib><creatorcontrib>Fredericks, Judith</creatorcontrib><creatorcontrib>Thompson, Kenneth D.</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of virological methods</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Baunoch, David A.</au><au>Lobell, Allegra</au><au>Kane, Dorothy</au><au>Fredericks, Judith</au><au>Thompson, Kenneth D.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>The development of an automated in situ assay for the detection of human cytomegalovirus in peripheral blood leukocytes</atitle><jtitle>Journal of virological methods</jtitle><addtitle>J Virol Methods</addtitle><date>1999-08-01</date><risdate>1999</risdate><volume>81</volume><issue>1</issue><spage>31</spage><epage>37</epage><pages>31-37</pages><issn>0166-0934</issn><eissn>1879-0984</eissn><coden>JVMEDH</coden><abstract>Human cytomegalovirus (HCMV) infections are common in immunosuppressed patients, especially transplant recipients and patients with AIDS. The utility of an automated in situ hybridization (ISH) assay for the rapid detection of HCMV immediate early mRNA was evaluated using cytospin (Shandon Lipshaw, Inc., Pittsburgh, PA) prepared leukocytes from peripheral blood samples. In this study, the detection of HCMV immediate early protein by immunofluorescent antibody staining of the standard shell vial assay was compared to the detection of HCMV immediate early mRNA in peripheral blood leukocytes using the automated ISH system. Of 135 specimens tested, eight specimens were positive using HCMV ISH compared to seven positive specimens using shell vial assay. Overall, HCMV ISH demonstrated 100% sensitivity and 99% specificity. Since the HCMV ISH assay requires minimal labor, and can be completed in less than 5 h, this method should be evaluated as a potential replacement for shell vial assay for the diagnosis of HCMV infection.</abstract><cop>London</cop><cop>Amsterdam</cop><cop>New York, NY</cop><pub>Elsevier B.V</pub><pmid>10488758</pmid><doi>10.1016/S0166-0934(99)00062-2</doi><tpages>7</tpages></addata></record> |
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| subjects | Biological and medical sciences Cytomegalovirus Cytomegalovirus - genetics Cytomegalovirus - isolation & purification Fibroblasts - chemistry Fibroblasts - virology Fundamental and applied biological sciences. Psychology Human cytomegalovirus Human viral diseases Humans Immediate early mRNA Immediate-Early Proteins - genetics In situ hybridization In Situ Hybridization - instrumentation In Situ Hybridization - methods Infectious diseases Leukocytes Leukocytes - chemistry Leukocytes - virology Medical sciences Microbiology RNA, Messenger - genetics RNA, Viral - genetics Techniques used in virology Viral diseases Viral diseases of the lymphoid tissue and the blood. Aids Virology |
| title | The development of an automated in situ assay for the detection of human cytomegalovirus in peripheral blood leukocytes |
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