Validation of an Indirect Immunofluorescence Assay and Commercial Q Fever Enzyme-Linked Immunosorbent Assay for Use in Macropods
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Date
2022-07
Open Access Location
Journal Title
Journal ISSN
Volume Title
Publisher
American Society for Microbiology
Rights
(c) 2022 The Author/s
CC BY 4.0
CC BY 4.0
Abstract
Kangaroos are considered to be an important reservoir of Q fever in Australia, although there is limited knowledge on the true prevalence and distribution of coxiellosis in Australian macropod populations. Serological tests serve as useful surveillance tools, but formal test validation is needed to be able to estimate true seroprevalence rates, and few tests have been validated to screen wildlife species for Q fever. In this study, we modified and optimized a phase-specific indirect immunofluorescence assay (IFA) for the detection of IgG antibodies against Coxiella burnetii in macropod sera. The assay was validated against the commercially available ID Screen Q fever indirect multispecies enzyme-linked immunosorbent assay (ELISA) kit (IDVet, Grabels, France) to estimate the diagnostic sensitivity and specificity of each assay, using Bayesian latent class analysis. A direct comparison of the two tests was performed by testing 303 serum samples from 10 macropod populations from the east coast of Australia and New Zealand. The analysis indicated that the IFA had relatively high diagnostic sensitivity (97.6% [95% credible interval [CrI], 88.0 to 99.9]) and diagnostic specificity (98.5% [95% CrI, 94.4 to 99.9]). In comparison, the ELISA had relatively poor diagnostic sensitivity (42.1% [95% CrI, 33.7 to 50.8]) and similar diagnostic specificity (99.2% [95% CrI, 96.4 to 100]) using the cutoff values recommended by the manufacturer. The estimated true seroprevalence of C. burnetii exposure in the macropod populations included in this study ranged from 0% in New Zealand and Victoria, Australia, up to 94.2% in one population from New South Wales, Australia.
Description
Keywords
Bayesian latent class models, Coxiella burnetii, ELISA, Q fever, enzyme-linked immunosorbent assay, immunofluorescence assay, macropods, sensitivity, specificity, test validation, Antibodies, Bacterial, Bayes Theorem, Coxiella burnetii, Enzyme-Linked Immunosorbent Assay, Fluorescent Antibody Technique, Indirect, Humans, Q Fever, Seroepidemiologic Studies, Victoria
Citation
Tolpinrud A, Stenos J, Chaber A-L, Devlin JM, Herbert C, Pas A, Dunowska M, Stevenson MA, Firestone SM. (2022). Validation of an Indirect Immunofluorescence Assay and Commercial Q Fever Enzyme-Linked Immunosorbent Assay for Use in Macropods.. J Clin Microbiol. 60. 7. (pp. e0023622-).