Evaluation of the Limulus amebocyte lysate and recombinant factor C assays for assessment of airborne endotoxin.

نویسندگان

  • Peter S Thorne
  • Sarah S Perry
  • Rena Saito
  • Patrick T O'Shaughnessy
  • John Mehaffy
  • Nervana Metwali
  • Thomas Keefe
  • Kelley J Donham
  • Stephen J Reynolds
چکیده

As a potent inflammatory agent, endotoxin is a key analyte of interest for studies of lung ailments in domestic environments and occupational settings with organic dust. A relatively unexplored advance in endotoxin exposure assessment is the use of recombinant factor C (rFC) from the Limulus pathway in a fluorometric assay. In this study, we compared airborne endotoxin concentrations in laboratory- and field-collected parallel air samples using the kinetic Limulus amebocyte lysate (LAL) assay and the rFC assay. Air sampling was performed using paired Institute of Occupational Medicine (IOM) samplers, Button samplers, closed-face cassettes, and cyclone samplers. Field sampling was performed in 10 livestock production facilities, including those housing swine, chicken, turkey, dairy cows, cattle, and horses. Laboratory sampling was performed in exposure chambers using resuspended airborne dust collected in five livestock facilities. Paired samples were extracted in pyrogen-free water with 0.05% Tween 20 and analyzed using LAL and rFC assays. In 402 field sample pairs there was excellent agreement between endotoxin concentrations determined by LAL and rFC (r = 0.93; P < 0.0001). In 510 laboratory sample pairs there was also excellent agreement between the two assays (r = 0.86; P < 0.0001). Correlations for subgroups of facility or dust type ranged from 0.65 to 0.96. Mixed-model analysis of variance (ANOVA) for the field studies showed significant interactions of facility-sampler and facility-assay. rFC/LAL ratios of the geometric means were 0.9 to 1.14 for the samplers (not significantly different from 1.0). The data from this study demonstrate that the LAL assay and the rFC assay return similar estimates of exposure in livestock facilities. Both methods provided suitable lower limits of detection such that all but 19 of 1,824 samples were quantifiable.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Endotoxin Contamination of Large Volume Parenterals as Detected by Limulus Amebocyte Lysate as an Alternative to Rabbit Pyrogen Test

The possibility of replacing the rabbit pyrogen test by the Limulus Amebocyte lysate (LAL) test, as a final release test for Large Volume Parenterals (LVPs) was investigated. The sensitivity and specificity of the LAL test and rabbit pyrogen assay were studied by means of artificially contaminated parenterals. Various LVPs (mostly containing electrolytes) were spiked with 0.5 and 5 EUlml o...

متن کامل

Measuring of free endotoxin in alum-precipitated vaccine of haemorrhagic septicaemia by limulus amebocyte lysate test

Haemorrhagic septicaemia (HS) vaccine which is prepared in Razi Institute is used in endemic areas ofIran. Aluminum-hydroxide gel was used as adjuvant for preparing this vaccine. Post-vaccinal shock reactionswere the main complaint after use of this vaccine. In a previous study, we could improve the vaccine byalum-precipitation Pasteurella multocida cells and removing the liquid phase. In this ...

متن کامل

Biological Activity of Masked Endotoxin

Low endotoxin recovery (LER) is a recently discovered phenomenon describing the inability of limulus amebocyte lysate (LAL)-based assays to detect lipopolysaccharide (LPS) because of a "masking effect" caused by chelators or detergents commonly used in buffer formulations for medical products and recombinant proteins. This study investigates the masking capacities of different buffer formulatio...

متن کامل

Airborne endotoxin from indoor and outdoor environments: effect of sample dilution on the kinetic Limulus amebocyte lysate (LAL) assay.

Airborne endotoxins in occupational environments are a potential respiratory hazard to individuals. In this study, airborne endotoxins were collected using open-face and button aerosol samplers from inside animal housing units and downwind from agricultural production sites and a wastewater treatment plant. Filter extracts were then diluted to examine the effect of interfering substances on the...

متن کامل

Biomedical Applications of Limulus Amebocyte Lysate

This year celebrates the 30th anniversary of the licensing of Limulus amebocyte lysate (LAL) by the US Food and Drug Administration (FDA) as a test for the presence of endotoxin in biologicals, pharmaceutical drugs, and medical devices. LAL is currently recognized by several major pharmacopoeias and is used worldwide. That a suitable alternative for the detection of endotoxin has not supplanted...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:
  • Applied and environmental microbiology

دوره 76 15  شماره 

صفحات  -

تاریخ انتشار 2010