Recombinant factor C assay for measuring endotoxin in house dust: comparison with LAL, and (1 --> 3)-beta-D-glucans

Am J Ind Med. 2006 Apr;49(4):296-300. doi: 10.1002/ajim.20264.

Abstract

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.

Conclusion: 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.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Air Microbiology
  • Arthropod Proteins
  • Dust / analysis*
  • Endotoxins / analysis*
  • Enzyme Precursors*
  • Enzyme-Linked Immunosorbent Assay
  • Humans
  • Limulus Test / methods*
  • Recombinant Proteins
  • Serine Endopeptidases*
  • beta-Glucans*

Substances

  • Arthropod Proteins
  • Dust
  • Endotoxins
  • Enzyme Precursors
  • Recombinant Proteins
  • beta-Glucans
  • Serine Endopeptidases
  • Limulus clotting factor C