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.
