Recombinant factor C assay for measuring endotoxin in house dust: comparison with LAL, and (1 --> 3)-beta-D-glucans.
BACKGROUND
Measurement of exposure to environmental endotoxin is frequently performed using a Limulus amebocyte lysate (LAL) based assay. Recently, a new method has become available with similar sensitivity and potentially greater specificity using recombinant Factor C (rFC) from the horseshoe crab Carcinoscorpius rotundicauda. A preliminary study was carried out to determine the comparability of LAL and rFC in measuring endotoxins in house dust for large scale epidemiologic studies.


METHODS
House dust samples were collected from family rooms by vacuuming 1 m2 of the center of the room. Sixty sieved house dust samples were assayed for endotoxin by LAL (Cambrex, KQCL lysate) and rFC (Pyrogene, Cambrex) and for (1 --> 3)-beta-D-glucans by ELISA. The resistant parallel line estimation was used for data analysis of LAL and rFC. A four-parameter logistic fit with inverse prediction was used to calculate (1 --> 3)-beta-D-glucan levels of the samples.


RESULTS
The spike recovery was 113.63% (95% CI = 101.69, 125.57%) for LAL and 99.69% (95% CI = 90.14, 109.24%) for rFC assays. The LAL assay gave higher endotoxin estimates compared with rFC. The LAL and rFC estimates were highly correlated (r = 0.86, P < 0.0001). The difference between LAL and rFC endotoxin estimates correlated with the LAL estimates (r = 0.51, P < 0.0001). However, the difference was not correlated with (1 --> 3)-beta-D-glucans.


CONCLUSIONS
LAL and rFC gave comparable results, hence either assay can be used for studies of endotoxin exposure. The current study shows that (1 --> 3)-beta-D-glucan is not a major factor interfering with endotoxin measurements in house dust using a Cambrex KQCL LAL preparation.
Alwis
K Udeni
KU
Department of Environmental Health, Exposure, Epidemiology and Risk Assessment Program, Harvard School of Public Health, Landmark Center West, Boston, Massachusetts, USA.
Milton
Donald K
DK
D000391:Air Microbiology; D060829:Arthropod Proteins; D004391:Dust / Q000032:analysis*; D004731:Endotoxins / Q000032:analysis*; D004792:Enzyme Precursors; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D008033:Limulus Test / Q000379:methods*; D011994:Recombinant Proteins; D012697:Serine Endopeptidases; D047071:beta-Glucans
D060829:Arthropod Proteins; D004391:Dust; D004731:Endotoxins; D004792:Enzyme Precursors; D011994:Recombinant Proteins; D047071:beta-Glucans; D012697:Serine Endopeptidases; C057151:Limulus clotting factor C