Development of a quantitative sandwich enzyme-linked immunosorbent assay for detecting the MPT64 antigen of mycobacterium tuberculosis

Mijung Ji, Byungki Cho, Young Shik Cho, Song Yong Park, Sang Nae Cho, Bo Young Jeon, Byoung Su Yoon

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8 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)746-752
Number of pages7
JournalYonsei medical journal
Volume55
Issue number3
DOIs
Publication statusPublished - 2014 May

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All Science Journal Classification (ASJC) codes

  • Medicine(all)

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