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Serodiagnosis of environmental mycobacterial infections
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
Affiliation:
  • a Cantacuzino Institute, Mycobacterial Antigens Department, Bucharest, Romania
  • b “Marius Nasta” Pneumophtisiology Institute, Bucharest, Romania
  • c “Dr. Victor Babes” Hospital for Tropical and Infectious Diseases, Bucharest, Romania
  • d “Carol Davila” Emergency Military Clinical Hospital, Bucharest, Romania
  • e “Carol Davila” University of Medicine and Pharmacy, Bucharest, Romania
  • f Colorado State University, Fort Collins, CO, USA
  • 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.
    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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