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Measurement of β-(1,3)-glucan in household dust samples using Limulus amebocyte assay and enzyme immunoassays: an
Collin R Brooks1, Rob Siebers, Julian Crane
1Centre for Public Health Research, Massey University Wellington Campus, Wellington, New Zealand. C.R.Brooks@massey.ac.nz.
Abstract:
Environmental levels of β-(1,3)-glucan, an inflammatory fungal cell wall component, have been suggested to be related to respiratory symptoms. However there is currently little data comparing β-(1,3)-glucan detection methods and/or results obtained in different laboratories. The aim of this study was to compare levels of β-(1,3)-glucans detected in household dust samples (n = 40) using different extraction/detection methods (Limulus amebocyte assay (LAL), inhibition enzyme immunoassay (EIA) and sandwich EIA) in five different laboratories. Dust sample aliquots were sent to participating centres, extracted and analysed for β-(1,3)-glucan according to standard in-house procedures. Significant differences in the levels of β-(1,3)-glucan were observed between all laboratories (geometric mean levels ranging from 15.4 μg g (-1) to 4754 μg g(-1) dust; p < 0.0001) with the exception of those using a similar LAL method. The inhibition EIA used in laboratory D produced mean β-(1,3)-glucan measurements 80-100 times higher than the LAL assays, 4 times higher than the sandwich EIA in the same lab, 17.6 times those obtained with the EIA in lab E and 363 times those obtained in the EIA in laboratory C. Pearson's correlations generally showed significant associations between methods and laboratories, particularly those using similar methodology (R ranging from 0.5 to 0.8; p < 0.001), although some poor and even inverse correlations were observed. Bland-Altman analyses showed moderate to good agreement between most assays, although clear absolute differences were observed. In conclusion, although results obtained with different methods were often significantly correlated and therefore comparable in relative terms, direct comparison of results between laboratories and assays may be inappropriate.
Insights
Comparing methods for detecting fungal beta-(1,3)-glucan in dust revealed significant laboratory variations. While some methods showed correlation, direct comparison of results between labs and assays may be inappropriate.
Area of Science:
- Environmental Science
- Mycology
- Analytical Chemistry
Background:
- Beta-(1,3)-glucan, a fungal cell wall component, is linked to respiratory issues.
- Limited data exists on comparing beta-(1,3)-glucan detection methods and inter-laboratory results.
Purpose of the Study:
- To compare beta-(1,3)-glucan levels in household dust using different extraction/detection methods.
- To assess inter-laboratory variability in beta-(1,3)-glucan measurements.
Main Methods:
- Forty household dust samples were analyzed by five laboratories.
- Methods included Limulus amebocyte assay (LAL), inhibition enzyme immunoassay (EIA), and sandwich EIA.
- Standard in-house extraction and analysis procedures were followed.
Main Results:
- Significant differences in beta-(1,3)-glucan levels were found between laboratories (p < 0.0001), except for similar LAL methods.
- Inhibition EIA yielded results 80-363 times higher than other methods.
- Pearson's correlations showed associations between similar methods, but Bland-Altman analyses revealed absolute differences.
Conclusions:
- While results from different methods can be comparable in relative terms, direct comparison is often inappropriate.
- Significant inter-laboratory variability exists for beta-(1,3)-glucan detection in dust.
- Standardization of methods is crucial for accurate environmental health assessments.
