Projects
Towards Standardization of Microparticle-Based, Solid Phase, HLA Antibody Identification Assays.
Robert Bray , Howard Gebel
Rationale:
Solid phase antibody detection assays have become a “standard” in many, if not most, clinical histocompatibility laboratories. Although very sensitive, recent studies have shown that not all donor-specific HLA antibodies (DSA) identified by the new technologies are predictive of a positive crossmatch or correlate with a poor clinical outcome. While there are many reasons to explain this lack of direct correlation, lack of standardization for such assays may be a significant contributor. Recent findings from a pilot proficiency testing survey, organized by our laboratory, involving laboratories in Canada and Australia, results demonstrated that a lack of standardization was a major factor contributing to discordant results between laboratories. In addition, inconsistencies between manufactures and between differing product lots from the same manufacture further compound the problem. Finally, calculation and interpretation of the data vary significantly between laboratories. Together, these variables result in data that are not comparable across laboratories. Hence, if we are to have some agreement as to the clinical relevance of antibodies detected by solid-phase methods, strategies are needed to standardize these assays.
Proposal:
In this workshop section we propose to develop standardized strategies and best practices for HLA antibody detection using microparticle-based, solid phase assays. Standardization strategies will included; a common methodological approach, common reagents (eg: secondary antibody); common negative control sera and most importantly, the development of a set of well characterized sera for use in subsequent comparisons by laboratories or vendors. Such well-characterized sera will provide not only a consensus for antibody specificity but a consensus for the strength (MFI or other fluorescence measurement) of the antibody such that laboratories can assess their ability to achieve comparable results. We propose to utilize sera that have already been reasonably characterized by our laboratory and by 15 laboratories in Canada via a proficiency testing program. We also propose to enroll a limited number of laboratories (max. 20-25) from varying geographic regions. In a manner similar to our Canadian/Australian testing experience, we will send well characterized sera to participating laboratories for evaluation. Laboratories may also be provided with some common reagents to use in addition to their own reagents as well as common testing protocol. Results will be reviewed and assessed for consensus. Following initial data review, a consensus set of data will be assembled for the sera. Laboratories will then be able to adjust protocols/reagents to achieve the consensus results. A final send out of new, but well characterized sera will be sent to determine laboratory performance. Finally, summary data will be presented at the International Workshop meeting to put forward standardized methods/protocols as well as provide some well characterized sera for laboratories and vendors to use.
Contact:
Robert Bray, PhD and Howard Gebel, PhD
Emory University Hospital,
Atlanta, GA 30322
USA
Email: rbray [at] emory [dot] edu.
Download a PDF of the ASHI 2010 Presentation - download PDF file.
Download a PDF of the ASHI 2011 Presentation - download PDF file.
