A new homogeneous assay for high throughput serological diagnosis of brucellosis in ruminants
| Autor: | John McGiven, Simon D. Brew, Jason Sawyer, Kate Harper, Alan Fisher, Stuart McLarnon, J. A. Stack, Nicola J. Commander, Lorraine L. Perrett |
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| Rok vydání: | 2008 |
| Předmět: |
Immunology
Brucella abortus Sheep Diseases Enzyme-Linked Immunosorbent Assay Brucella Biology Cross Reactions Sensitivity and Specificity Brucellosis Serology Brucellosis Bovine Antibody Specificity Predictive Value of Tests medicine Immunology and Allergy Animals False Positive Reactions Serologic Tests Throughput (business) Yersinia enterocolitica Goat Diseases Sheep Goats Reproducibility of Results medicine.disease biology.organism_classification Serum samples Virology Antibodies Bacterial Low volume Specific antibody Homogeneous Cattle |
| Zdroj: | Journal of immunological methods. 337(1) |
| ISSN: | 0022-1759 |
| Popis: | The control and eradication of brucellosis is highly desirable but heavily resource intensive as high throughput serological testing is required. The aim of this study was to meet the needs of high throughput screening laboratories involved in this process through the development of a new assay. An existing cELISA used for the serodiagnosis of brucellosis in ruminants was converted to an AlphaLISA homogenous proximity based assay. This assay requires no separation steps and can be performed in low volume microtitre format. The Brucella AlphaLISA was validated on a panel of bovine, ovine and caprine sera from infected and uninfected animals. The diagnostic sensitivities (>96%) and specificities (>98%) obtained compared well to those from cELISA, iELISA and FPA performed on the same samples. The AlphaLISA met the testing criteria set for ELISAs as defined by the OIEELISA standards and had an analytical sensitivity similar to that of the parent cELISA. The method was also used on a small panel of serum samples from cattle that were experimentally infected with Yersinia enterocolitica O:9. Some false positive reactions were obtained as was also the case with results from FPA, iELISA, cELISA, CFT and SAT. Despite this, the methodological advantages of the AlphaLISA mean that this assay is well suited to high throughput serodiagnosis. This report is the first description of the use of AlphaLISA to detect pathogen specific antibodies. Furthermore, the relative ease with which the cELISA was converted to this platform indicates that this technology is ready to meet the high throughput testing requirements for the diagnosis of many other diseases. |
| Databáze: | OpenAIRE |
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