원문정보
초록
영어
LAL (Limulus amebocyte lysate) test is used to detect bacterial endotoxin or LPS (lipopolysaccharide), a membrane component, of gram negative bacteria. There are 3
methods available for conventional testing: gel clotting, KTA (kinetic turbidimetric assay) and KCA (kinetic chromogenic assay). These require long incubation and expensive reagents. And LOC (lab-on-a-chip) is one of the solutions. We evaluated the assays' feasibility in terms of reducing assay time and volume. Among the 3, gel clotting was excluded because its result was too rough to be quantitative. The volume of LAL and CSE (control standard endotoxin) mixture was from 50 to 200 μl, and its final concentration was from 0.05 to 5 EU/ml. The test was triplicated. 96-well plate reader read the absorbance of a well every 3 min during 60±1 min at 340 nm for KTA,
and 405 nm for KCA with 37±1°C. Absorbance per light path length of each reaction volume was calculated. Absorbance controlled regression analysis showed the coefficient of determination above 0.98. These data suggested the assay time was shortened by 1/2 hr than the conventional methods and the LOC could distinguish various concentration of samples.