Highly quantitative serological detection of anti-cytomegalovirus (CMV) antibodies
<p>Abstract</p> <p>Background</p> <p>Human cytomegalovirus infection is associated with a variety of pathological conditions including retinitis, pneumonia, hepatitis and encephalitis that may be transmitted congenitally, horizontally and parenterally and occurs both as...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2009-05-01
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| Series: | Virology Journal |
| Online Access: | http://www.virologyj.com/content/6/1/45 |
| _version_ | 1818236282874101760 |
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| author | Alter Harvey J Drew W Lawrence Exner Maurice Ching Kathryn H Issa Alexandra T Burbelo Peter D Iadarola Michael J |
| author_facet | Alter Harvey J Drew W Lawrence Exner Maurice Ching Kathryn H Issa Alexandra T Burbelo Peter D Iadarola Michael J |
| author_sort | Alter Harvey J |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>Human cytomegalovirus infection is associated with a variety of pathological conditions including retinitis, pneumonia, hepatitis and encephalitis that may be transmitted congenitally, horizontally and parenterally and occurs both as a primary infection and as reactivation in immunocompromised individuals. Currently, there is a need for improved quantitative serological tests to document seropositivity with high sensitivity and specificity.</p> <p>Methods</p> <p>Here we investigated whether luciferase immunoprecipitation systems (LIPS) would provide a more quantitative and sensitive method for detecting anti-CMV antibodies. Four protein fragments of immunodominant regions of CMV antigens pp150 and pp65 were generated as <it>Renilla </it>luciferase (Ruc) fusion proteins and used in LIPS with two cohorts of CMV positive and negative sera samples previously tested by ELISA.</p> <p>Results</p> <p>Analysis of the antibody responses to two of these antigen fragments, pp150-d1 and pp150-d2, revealed geometric mean antibody titers in the first cohort that were 100–1000 fold higher in the CMV positive sera compared to the CMV negative samples (p < 0.0001) and infection status exactly matched the ELISA results for the 46 samples of the first cohort (100% sensitivity and 100% specificity). Two additional antigen fragments, pp65-d1 and pp65-d2 also showed robust antibody titers in some CMV-infected sera and yielded 50% and 96% sensitivity, respectively. Analysis of a second cohort of 70 samples using a mixture of the 4 antigens, which simplifies data collection and analysis, yielded values which correlated well with the sum of the values from the 4 separate tests (<it>r</it><sub><it>s </it></sub>= 0.93, p < 0.00001). While comparison of the LIPS results from this second cohort with ELISA showed 100% sensitivity, LIPS detected six additional CMV positive samples that were not detected by ELISA. Heat map analysis revealed that several of the LIPS positive/ELISA negative samples had positive LIPS immunoreactivity with 3–4 of the CMV antigens.</p> <p>Conclusion</p> <p>These results suggest that LIPS provides a highly robust and quantitative method for studying anti-CMV antibodies and has the potential to more accurately document CMV infection than standard ELISA.</p> |
| first_indexed | 2024-12-12T12:07:24Z |
| format | Article |
| id | doaj.art-969990d546974b44824b61a9f9b24e7c |
| institution | Directory Open Access Journal |
| issn | 1743-422X |
| language | English |
| last_indexed | 2024-12-12T12:07:24Z |
| publishDate | 2009-05-01 |
| publisher | BMC |
| record_format | Article |
| series | Virology Journal |
| spelling | doaj.art-969990d546974b44824b61a9f9b24e7c2022-12-22T00:24:58ZengBMCVirology Journal1743-422X2009-05-01614510.1186/1743-422X-6-45Highly quantitative serological detection of anti-cytomegalovirus (CMV) antibodiesAlter Harvey JDrew W LawrenceExner MauriceChing Kathryn HIssa Alexandra TBurbelo Peter DIadarola Michael J<p>Abstract</p> <p>Background</p> <p>Human cytomegalovirus infection is associated with a variety of pathological conditions including retinitis, pneumonia, hepatitis and encephalitis that may be transmitted congenitally, horizontally and parenterally and occurs both as a primary infection and as reactivation in immunocompromised individuals. Currently, there is a need for improved quantitative serological tests to document seropositivity with high sensitivity and specificity.</p> <p>Methods</p> <p>Here we investigated whether luciferase immunoprecipitation systems (LIPS) would provide a more quantitative and sensitive method for detecting anti-CMV antibodies. Four protein fragments of immunodominant regions of CMV antigens pp150 and pp65 were generated as <it>Renilla </it>luciferase (Ruc) fusion proteins and used in LIPS with two cohorts of CMV positive and negative sera samples previously tested by ELISA.</p> <p>Results</p> <p>Analysis of the antibody responses to two of these antigen fragments, pp150-d1 and pp150-d2, revealed geometric mean antibody titers in the first cohort that were 100–1000 fold higher in the CMV positive sera compared to the CMV negative samples (p < 0.0001) and infection status exactly matched the ELISA results for the 46 samples of the first cohort (100% sensitivity and 100% specificity). Two additional antigen fragments, pp65-d1 and pp65-d2 also showed robust antibody titers in some CMV-infected sera and yielded 50% and 96% sensitivity, respectively. Analysis of a second cohort of 70 samples using a mixture of the 4 antigens, which simplifies data collection and analysis, yielded values which correlated well with the sum of the values from the 4 separate tests (<it>r</it><sub><it>s </it></sub>= 0.93, p < 0.00001). While comparison of the LIPS results from this second cohort with ELISA showed 100% sensitivity, LIPS detected six additional CMV positive samples that were not detected by ELISA. Heat map analysis revealed that several of the LIPS positive/ELISA negative samples had positive LIPS immunoreactivity with 3–4 of the CMV antigens.</p> <p>Conclusion</p> <p>These results suggest that LIPS provides a highly robust and quantitative method for studying anti-CMV antibodies and has the potential to more accurately document CMV infection than standard ELISA.</p>http://www.virologyj.com/content/6/1/45 |
| spellingShingle | Alter Harvey J Drew W Lawrence Exner Maurice Ching Kathryn H Issa Alexandra T Burbelo Peter D Iadarola Michael J Highly quantitative serological detection of anti-cytomegalovirus (CMV) antibodies Virology Journal |
| title | Highly quantitative serological detection of anti-cytomegalovirus (CMV) antibodies |
| title_full | Highly quantitative serological detection of anti-cytomegalovirus (CMV) antibodies |
| title_fullStr | Highly quantitative serological detection of anti-cytomegalovirus (CMV) antibodies |
| title_full_unstemmed | Highly quantitative serological detection of anti-cytomegalovirus (CMV) antibodies |
| title_short | Highly quantitative serological detection of anti-cytomegalovirus (CMV) antibodies |
| title_sort | highly quantitative serological detection of anti cytomegalovirus cmv antibodies |
| url | http://www.virologyj.com/content/6/1/45 |
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