Serodiagnosis of environmental mycobacterial infections |
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Authors: | Stavri Henriette Ulea Irina Radu Dorel L Branaru Manuela Gheorghiu Moldovan Olga Bogdan Miron A Tudose Cornelia Raileanu Marinela Duiculescu Dan Ene Luminita Olar Viorel Ionita Catalin Popa Gabriela Loredana Popa Mircea I Brennan Patrick J |
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Affiliation: | a Cantacuzino Institute, Mycobacterial Antigens Department, Bucharest, Romaniab “Marius Nasta” Pneumophtisiology Institute, Bucharest, Romaniac “Dr. Victor Babes” Hospital for Tropical and Infectious Diseases, Bucharest, Romaniad “Carol Davila” Emergency Military Clinical Hospital, Bucharest, Romaniae “Carol Davila” University of Medicine and Pharmacy, Bucharest, Romaniaf Colorado State University, Fort Collins, CO, USA |
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Abstract: | To demonstrate the usefulness of enzyme-linked immunosorbent assay for serodiagnosis of mycobacterioses due to environmental mycobacteria we utilized a panel of glycolipid antigens selective for Mycobacterium avium-intracellulare, Mycobacterium kansasii, Mycobacterium xenopi, Mycobacterium scrofulaceum and Mycobacterium gordonae. The levels of circulating antibodies were determined against the environmental mycobacteria, and Mycobacterium tuberculosis in human immunodeficiency virus-negative and -positive patient sera. The method used immunomagnetic separation of the antigens, with covalent immobilization of antibodies to superparamagnetic amine and carboxyl terminated particles in solutions of the specific antigens. Enzyme-linked immunosorbent assay was performed on 195 patient sera: 34 with infections due to environmental mycobacteria, 114 with tuberculosis, 47 with other respiratory diseases. There were 46 human immunodeficiency virus-1 infected individuals. Among the 34 infections due to environmental mycobacteria, 9 patients were singularly infected with an environmental mycobacterium, and 25 co-infected with both M. tuberculosis and an environmental mycobacterium. Sensitivity, specificity and false positivity ranges were determined for each of the volunteer groups: tuberculosis positive, human immunodeficiency virus negative; tuberculosis positive, human immunodeficiency virus positive; those with infections due to individual environmental mycobacteria (such as M. scrofulaceum and M. kansasii); and those with other respiratory diseases. We demonstrate that such multiple assays, can be useful for the early diagnosis of diverse environmental mycobacterial infections to allow the start of treatment earlier than henceforth. |
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Keywords: | EM, environmental mycobacteria M. tuberculosis, Mycobacterium tuberculosis TB, tuberculosis AFB, acid-fast bacillus ELISA, enzyme-linked immunosorbent assay GPL, glycopeptidolipid LOS, lipooligosaccharides PGL, phenolic glycolipids MAI, M. avium/M.intracellulare ATS, American Thoracic Society NTM, non-tuberculous mycobacteria ORD, other respiratory diseases GL, glycolipid TLC, thin layer chromatography PBS, phosphate buffered saline IgG, immunoglobulins G BSA, Bovine Serum Albumin MES, 2-(N-morpholino)ethanesulfonic acid MP, magnetic particles C18, MPG CPG-C18, (magnetic porous glass particles, with a lateral chain of 18 Carbon atoms BM, BioMag® superparamagnetic Iron Oxide HRP- SpA, Horseradish peroxidase- Staphylococcal protein A PPV, positive predictive value NPV, negative predictive value FNR, false negative rate FPR, false positive rate GL-NH2, GL-BioMag ® Plus Amine- M. tuberculosis H37Rv IgG GL-COOH, GL-BioMag ® Plus Carboxyl- M. tuberculosis H37Rv IgG OD, optical density IU, immunoenzymatic units HIV, human immunodeficiency virus |
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