title: Impact of sensitivity of human leucocyte antigen antibody detection by Luminex technology on graft loss at 1 year.
abstract: BACKGROUND
  The clinical relevance of the detection of human leucocyte antigen (HLA) antibodies in sera of renal transplant recipients by highly sensitive methods such as Luminex alone is uncertain and a matter of debate. The choice of output thresholds affects antibody detection and thus organ allocation, yet there are no internationally agreed threshold levels. This study aims at evaluating our current practice of using an MFI threshold of 1000 in antibody detection.
  
  
  METHODS
  We carried out a case-control study by looking at 761 renal transplant recipients at one unit between 2000 and 2010. Of these, there were 93 cases of graft loss within 1 year and stored serum samples of 40 cases were available for testing. Controls were selected (graft function >2 years) and individually matched according to age, sex, number of transplants and date of transplant. All 40 cases and 40 controls had negative crossmatch by complement-dependent cytotoxicity (CDC) at the time of transplant, and pre-transplant sera were re-analysed for the presence of detectable HLA and donor-specific antibodies (DSAs) using Luminex screen and single-antigen beads and MFI threshold values of 1000, 2000 and 4000.
  
  
  RESULTS
  In nearly 48% of cases with graft loss within a year, HLA antibodies were detectable by Luminex when using a 1000 MFI threshold. This was 25% greater than in controls (P = 0.017). There was also a 15% increase in detected DSAs; however, statistical significance depends on the inclusion or exclusion of one specific case. Using MFI thresholds of 2000 and 4000, no DSAs were found in any long-term surviving grafts.
  
  
  CONCLUSIONS
  Selection of appropriate MFI cut-off values influences the detection of DSAs and, thus, organ allocation. Using a threshold of 1000 led to the detection of DSAs in 5% of long-term graft survivors in our population and should be considered too sensitive. Using a detection threshold of 2000 is sufficiently sensitive and leads to clinically relevant detection of DSA.
authors.affiliation: Department of Transplant Surgery , Royal Liverpool University Hospital , Liverpool , UK.
authors.forename: Peter
authors.initials: P
authors.lastname: Szatmary
authors.affiliation: Department of Transplant Surgery , Royal Liverpool University Hospital , Liverpool , UK.
authors.forename: James
authors.initials: J
authors.lastname: Jones
authors.affiliation: Department of Transplant Surgery , Royal Liverpool University Hospital , Liverpool , UK.
authors.forename: Abdul
authors.initials: A
authors.lastname: Hammad
authors.affiliation: Department of Transplant Surgery , Royal Liverpool University Hospital , Liverpool , UK.
authors.forename: Derek
authors.initials: D
authors.lastname: Middleton
keywords: HLA; Luminex; crossmatch; renal transplantation