Z Gastroenterol 2013; 51 - P_1_58
DOI: 10.1055/s-0032-1331958

Evaluation of an automated hyaluronan latex agglutination assay enables the implementation of routine laboratory HEPASCORE calculation

E Yagmur 1, M Haumann 1, A Koch 2, H Lutz 2, C Trautwein 2, F Tacke 2
  • 1Medical Care Center, Dr. Stein and colleagues, Laboratory medicine, Mönchengladbach, Germany
  • 2RWTH-University Hospital Aachen, Department of Medicine III, Aachen, Germany

Background: The non-invasive HEPASCORE was reported as a diagnostic tool for predicting liver fibrosis. To date, hyaluronan (HA) is still predominantly assayed by time-consuming and costly enzyme-linked protein binding assay (ELISA).

Methods: We aimed to evaluate the analytical performance of a novel automated HA-latex agglutination assay (WAKO, Osaka, Japan) using KoneLab 30i analyzer (ThermoFisher Scientific, Hudson, New Hampshire, USA) and compared HA-concentrations in healthy blood donors (N=57) to the reference HEPASCORE-ELISA (HA Test Kit, Corgenix, Westminster, USA).

Results: The lowest detectable level of automated HA was found to be 34ng/mL (95. percentile). Serial dilution of recombinant HA concentrations provided linear response within a dilution range from 9:10 to 1:10 (1000ng/mL, P<0.0001 and 100ng/mL, P<0.0001). The recovery of HA was 94%-114% (1000ng/mL) and 92%-130% (100ng/mL), respectively. To run the intra-assay precision (20 replicates/day), 2 recombinant HA concentrations (100 and 400ng/mL) and 1 serum pool (170ng/mL) were prepared. The intra-assay precision study resulted in total coefficients of variation (CV) of 5.6%-6.1% for recombinant HA concentrations and 7.9% for serum pool. Inter-assay-precision yielded CVs of 2.2% and 3.9% (recombinant HA concentrations 100 and 400ng/mL, respectively; 1 determination/day for 20 days). Comparison of HA concentrations in healthy blood donors measured with the automated latex agglutination assay and the reference HEPASCORE HA-ELISA showed significant correlation (r=0.8525; P<0.0001).

Conclusion: Our data indicate that automated HA-latex agglutination assay presents excellent analytical performance and satisfactory correlation compared to reference method. It is easy to perform, accurate and enables the implementation of an automated HEPASCORE as routinely available non-invasive index of liver fibrosis.