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J Microbiol Methods ; 127: 236-241, 2016 08.
Artigo em Inglês | MEDLINE | ID: mdl-27317896

RESUMO

Campylobacter enteritis in humans is primarily associated with C. jejuni/coli infection. The impact of other Campylobacter spp. is likely to be underestimated due to the bias of culture methods towards Campylobacter jejuni/coli diagnosis. Stool antigen tests are becoming increasingly popular and appear generally less species-specific. A review of independent studies of the ProSpecT® Campylobacter Microplate enzyme immunoassay (EIA) developed for C. jejuni/coli showed comparable diagnostic results to culture methods but the examination of non-jejuni/coli Campylobacter spp. was limited and the limit-of-detection (LOD), where reported, varied between studies. This study investigated LOD of EIA for Campylobacter upsaliensis, Campylobacter hyointestinalis and Campylobacter helveticus spiked in human stools. Multiple stools and Campylobacter isolates were used in three different concentrations (10(4)-10(9)CFU/ml) to reflect sample heterogeneity. All Campylobacter species evaluated were detectable by EIA. Multivariate analysis showed LOD varied between Campylobacter spp. and faecal consistency as fixed effects and individual faecal samples as random effects. EIA showed excellent performance in replicate testing for both within and between batches of reagents, in agreement between visual and spectrophotometric reading of results, and returned no discordance between the bacterial concentrations within independent dilution test runs (positive results with lower but not higher concentrations). This study shows how limitations in experimental procedures lead to an overestimation of consistency and uniformity of LOD for EIA that may not hold under routine use in diagnostic laboratories. Benefits and limitations for clinical practice and the influence on estimates of performance characteristics from detection of multiple Campylobacter spp. by EIA are discussed.


Assuntos
Campylobacter coli/isolamento & purificação , Fezes/microbiologia , Técnicas Imunoenzimáticas , Limite de Detecção , Carga Bacteriana , Infecções por Campylobacter/diagnóstico , Infecções por Campylobacter/microbiologia , Campylobacter coli/enzimologia , Campylobacter coli/imunologia , Campylobacter jejuni/enzimologia , Campylobacter jejuni/imunologia , Campylobacter jejuni/isolamento & purificação , Ensaio de Imunoadsorção Enzimática , Humanos , Reação em Cadeia da Polimerase , Sensibilidade e Especificidade
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