Development and validation of a novel diagnostic test for human brucellosis using a glyco-engineered antigen coupled to magnetic beads

Autor: Jeremy A. Iwashkiw, Mario F. Feldman, Jorge C. Wallach, Diego J. Comerci, Luciano J. Melli, Bettina Deodato, Juan E. Ugalde, Andres Eduardo Ciocchini, Diego A. Rey Serantes
Jazyk: angličtina
Rok vydání: 2013
Předmět:
Ciencias de la Salud
law.invention
purl.org/becyt/ford/1 [https]
Ciencias Naturales y Exactas
law
Tecnologías Que Involucran la Identificación de Adn
Proteínas y Enzimas
y Cómo Influyen En El Conjunto de Enfermedades y Mantenimiento del Bienestar
purl.org/becyt/ford/3.4 [https]
Yersinia enterocolitica
Ciencias Médicas y de la Salud
Immunoassay
0303 health sciences
medicine.diagnostic_test
biology
lcsh:Public aspects of medicine
Zoonosis
Bioquímica y Biología Molecular (ídem 3.1.10)
Antibodies
Bacterial
Microspheres
3. Good health
Infectious Diseases
Veterinary Diseases
Recombinant DNA
Medicine
purl.org/becyt/ford/3 [https]
Research Article
Biotechnology
lcsh:Arctic medicine. Tropical medicine
lcsh:RC955-962
Brucella
DIAGNOSIS
Sensitivity and Specificity
Microbiology
Campylobacter jejuni
Brucellosis
Biotecnología de la Salud
Ciencias Biológicas
Magnetics
03 medical and health sciences
purl.org/becyt/ford/3.3 [https]
Antigen
Diagnostic Medicine
medicine
Humans
BRUCELLA
purl.org/becyt/ford/1.6 [https]
Biology
030304 developmental biology
Antigens
Bacterial
Diagnostic Tests
Routine
030306 microbiology
business.industry
Public Health
Environmental and Occupational Health
RECOMBINANT GLYCOCONJUGATE
lcsh:RA1-1270
biology.organism_classification
medicine.disease
Virology
Enfermedades Infecciosas
HUMAN BRUCELLOSIS
Veterinary Science
business
Zdroj: PLoS Neglected Tropical Diseases, Vol 7, Iss 2, p e2048 (2013)
CONICET Digital (CONICET)
Consejo Nacional de Investigaciones Científicas y Técnicas
instacron:CONICET
PLoS Neglected Tropical Diseases
ISSN: 1935-2735
1935-2727
Popis: Brucellosis is a highly contagious zoonosis and still a major human health problem in endemic areas of the world. Although several diagnostic tools are available, most of them are difficult to implement especially in developing countries where complex health facilities are limited. Taking advantage of the identical structure and composition of the Brucella spp. and Yersinia enterocolitica O:9 O-polysaccharide, we explored the application of a recombinant Y. enterocolitica O:9-polysaccharide-protein conjugate (OAg-AcrA) as a novel antigen for diagnosis of human brucellosis. We have developed and validated an indirect immunoassay using OAg-AcrA coupled to magnetic beads. OAg-AcrA was produced and purified with high yields in Y. enterocolitica O:9 cells co-expressing the oligosaccharyltransferase PglB and the protein acceptor AcrA of Campylobacter jejuni without the need for culturing Brucella. Expression of PglB and AcrA in Y. enterocolitica resulted in the transfer of the host O-polysaccharide from its lipid carrier to AcrA. To validate the assay and determine the cutoff values, a receiver-operating characteristic analysis was performed using a panel of characterized serum samples obtained from healthy individuals and patients of different clinical groups. Our results indicate that, using this assay, it is possible to detect infection caused by the three main human brucellosis agents (B. abortus, B. melitensis and B. suis) and select different cutoff points to adjust sensitivity and specificity levels as needed. A cutoff value of 13.20% gave a sensitivity of 100% and a specificity of 98.57%, and a cutoff value of 16.15% resulted in a test sensitivity and specificity of 93.48% and 100%, respectively. The high diagnostic accuracy, low cost, reduced assay time and simplicity of this new glycoconjugate-magnetic beads assay makes it an attractive diagnostic tool for using not only in clinics and brucellosis reference laboratories but also in locations with limited laboratory infrastructure and/or minimally trained community health workers.
Author Summary Brucellosis is one of the most widespread zoonoses in the world and still represents a mayor animal and human health problem in many endemic areas. Central for early diagnosis and appropriate treatment of the disease in humans is to have an easy to implement, fast and accurate diagnostic test for the disease. Currently, most of the diagnostic tests for human brucellosis are not of simple implementation in areas where laboratory infrastructure or trained personnel are not available. We present here a novel and simple diagnostic test for human brucellosis using a glyco-engineered antigen coupled to magnetic beads. Our results show that, with this new method, it is possible to detect infection caused by the three main human brucellosis agents with high sensitivity and specificity. We additionally demonstrate that this assay allows the selection of different cutoff points so that the desired operating characteristics of the test in terms of diagnostic sensitivity and specificity can be adjusted according to the needs of the operator. We believe that the high accuracy, low cost and simplicity of this new test makes it an attractive diagnostic tool not only for clinics and brucellosis reference laboratories but also for locations with limited laboratory infrastructure and/or minimally trained community health workers.
Databáze: OpenAIRE
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