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Clinical and Vaccine Immunology, September 2009, p. 1327-1331, Vol. 16, No. 9
1071-412X/09/$08.00+0     doi:10.1128/CVI.00047-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Development and Validation of a Fluorescent Microsphere Immunoassay for Soluble CD30 Testing{triangledown}

Igor Pavlov,1* Thomas B. Martins,1 and Julio C. Delgado1,2

ARUP Institute for Clinical and Experimental Pathology,1 Department of Pathology, University of Utah School of Medicine, Salt Lake City, Utah2

Received 3 February 2009/ Returned for modification 23 March 2009/ Accepted 7 July 2009

Testing for soluble CD30 (sCD30), an indicator of Th2 immune response, is a useful prognostic marker in solid organ transplantation, lymphoproliferative disorders, autoimmunity, and various parasitic diseases. In this study we report the development and validation of a fluorescent microsphere immunoassay for the detection of sCD30 in serum, plasma, and culture supernatants. The dynamic range of this assay is 1 to 400 ng/ml, and the rate of recovery of various concentrations of recombinant sCD30 ranges from 97 to 116% (average recovery, 105%). The test showed a high degree of precision in both intra-assay and interassay studies (coefficients of variation, as high as 7% and 8%, respectively), with a sensitivity of 1 ng/ml. The normal reference range calculated for a cohort of 151 healthy individuals was 1 to 29 ng/ml. The clinical usefulness of the sCD30 fluorescent microsphere immunoassay was demonstrated by showing that levels of sCD30 have a positive correlation with specimens containing high titers of anti-double-stranded DNA antibodies and high titers of immunoglobulin G against Leishmania species. Given the multiplexing potential of the sCD30 fluorescent microsphere immunoassay reported in this study, it is expected that testing of sCD30 concentrations along with those of other cytokines will become an important diagnostic tool for selected immunological and inflammatory diseases where Th2-type cytokine responses have been reported.

* Corresponding author. Mailing address: ARUP Institute for Clinical and Experimental Pathology, 500 Chipeta Way, Salt Lake City, UT 84108. Phone: (801) 583-2787, ext. 2967. Fax: (801) 584-5048. E-mail: igor.pavlov{at}aruplab.com

{triangledown} Published ahead of print on 15 July 2009.

Clinical and Vaccine Immunology, September 2009, p. 1327-1331, Vol. 16, No. 9
1071-412X/09/$08.00+0     doi:10.1128/CVI.00047-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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