Development of a quantitative sandwich enzyme-linked immunosorbent assay for detecting the MPT64 antigen of Mycobacterium tuberculosis
| Autor: | Song Yong Park, Bo Young Jeon, Mijung Ji, Byoung Su Yoon, Young Shik Cho, Sang Nae Cho, Byungki Cho |
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| Rok vydání: | 2013 |
| Předmět: |
Tuberculosis
medicine.drug_class Enzyme-Linked Immunosorbent Assay medicine.disease_cause Monoclonal antibody Cross-reactivity law.invention Mycobacterium tuberculosis Antigen law sandwich ELISA Medicine MPT64 protein chemistry.chemical_classification Antigens Bacterial biology business.industry General Medicine biology.organism_classification medicine.disease Virology Highly sensitive Enzyme Infectious Diseases chemistry Recombinant DNA Original Article business |
| Zdroj: | Yonsei Medical Journal |
| ISSN: | 1976-2437 |
| Popis: | PURPOSE Tuberculosis (TB) is a major infectious disease and is responsible for two million deaths annually. For the identification and quantitation of Mycobacterium tuberculosis (M. tuberculosis), a causative agent of TB, a sandwich enzyme-linked immunosorbent assay (ELISA) against the MPT64 protein of M. tuberculosis, an antigen marker of the M. tuberculosis complex, was developed. MATERIALS AND METHODS The MPT64 protein was expressed, and anti-MPT64 monoclonal antibodies were prepared. A sandwich ELISA was established using recombinant MPT64 protein and anti-MPT64 monoclonal antibodies. The sandwich MPT64 ELISA was evaluated using reference and clinical mycobacterial strains. RESULTS The sandwich MPT64 ELISA detected MPT64 protein from 2.1 ng/mL to 250 ng/mL (equivalent to 1.7×10⁴ CFU/mL and 2.0×10⁶ CFU/mL). All 389 clinical M. tuberculosis isolates tested positive in the sandwich MPT64 ELISA (sensitivity, 100%), and the assay showed no cross reactivity to any tested nontuberculous mycobacterial strain (specificity, 100%). CONCLUSION The sandwich MPT64 ELISA is a highly sensitive and quantitative test for MPT64 protein, which can identify M. tuberculosis. |
| Databáze: | OpenAIRE |
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