Antigens secreted from Mycobacterium tuberculosis: Identification by proteomics approach and test for diagnostic marker

Yil Bahk Young, Am Kim Suk, Ji Soo Kim, Hyung Jin Euh, Gil Han Bai, Sang Nae Cho, Sam Kim Yu

Research output: Contribution to journalArticle

100 Citations (Scopus)

Abstract

Tuberculosis caused by mycobacteria, mainly Mycobacterium tuberculosis, is a major infectious disease of the respiratory system. An early diagnosis followed by chemotherapy is the major control strategy. In an effort to identify the antigens suitable for immunodiagnosis and vaccines, the proteins secreted in a culture medium from the M. tuberculosis K-strain, which is the most prevalent among the clinical isolates in Korea and belongs to the Beijing family, were analyzed by two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) and compared with those from the M. tuberculosis H37Rv and CDC1551 strains. Eight proteins, Rv0652, Rv1636, Rv2818c, Rv3369, Rv3865, Rv0566c, MT3304, and Rv3160, were identified by matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS) or liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) and found to be relatively abundant in the culture medium from the M. tuberculosis K-strain but less so from the CDC1551 or H37Rv strains. In addition, Rv3874 (CFP-10), Rv-0560c and Rv3648c, which were expressed increasingly in the K and CDC1551 strains, were also identified using the same proteomics technology. All proteins were prepared by molecular cloning, expression in Escherichia coli followed by affinity purification. Among them, three proteins, rRv3369, rRv0566c, and rRv3874, were selected by prescreening and examined fortheir potential as serodiagnostic antigens using an enzyme-linked immunosorbent assay. When 100 sera from tuberculosis patients and 100 sera from the healthy controls were analyzed, rRV3369, rRv3874, and rRv0566c showed a sensitivity of 60%, 74%, and 43%, and a specificity of 96%, 97%, and 84%, respectively. These results suggest that the rRv3369 and rRv3874 proteins, which were expressed more abundantly in the more recently obtained clinical isolates of M. tuberculosis than in the laboratory-adapted H37Rv strain, are promising for use in the serodiagnosis of tuberculosis.

Original languageEnglish
Pages (from-to)3299-3307
Number of pages9
JournalProteomics
Volume4
Issue number11
DOIs
Publication statusPublished - 2004 Nov 1

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Mycobacterium tuberculosis
Routine Diagnostic Tests
Proteomics
Antigens
Proteins
Mass spectrometry
Culture Media
Tuberculosis
Respiratory system
Electrospray ionization
Immunosorbents
Immunologic Tests
Chemotherapy
Electrospray Ionization Mass Spectrometry
Cloning
Liquid chromatography
Electrophoresis, Gel, Two-Dimensional
Molecular Cloning
Serologic Tests
Korea

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

Cite this

Young, Yil Bahk ; Suk, Am Kim ; Kim, Ji Soo ; Euh, Hyung Jin ; Bai, Gil Han ; Cho, Sang Nae ; Yu, Sam Kim. / Antigens secreted from Mycobacterium tuberculosis : Identification by proteomics approach and test for diagnostic marker. In: Proteomics. 2004 ; Vol. 4, No. 11. pp. 3299-3307.
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abstract = "Tuberculosis caused by mycobacteria, mainly Mycobacterium tuberculosis, is a major infectious disease of the respiratory system. An early diagnosis followed by chemotherapy is the major control strategy. In an effort to identify the antigens suitable for immunodiagnosis and vaccines, the proteins secreted in a culture medium from the M. tuberculosis K-strain, which is the most prevalent among the clinical isolates in Korea and belongs to the Beijing family, were analyzed by two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) and compared with those from the M. tuberculosis H37Rv and CDC1551 strains. Eight proteins, Rv0652, Rv1636, Rv2818c, Rv3369, Rv3865, Rv0566c, MT3304, and Rv3160, were identified by matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS) or liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) and found to be relatively abundant in the culture medium from the M. tuberculosis K-strain but less so from the CDC1551 or H37Rv strains. In addition, Rv3874 (CFP-10), Rv-0560c and Rv3648c, which were expressed increasingly in the K and CDC1551 strains, were also identified using the same proteomics technology. All proteins were prepared by molecular cloning, expression in Escherichia coli followed by affinity purification. Among them, three proteins, rRv3369, rRv0566c, and rRv3874, were selected by prescreening and examined fortheir potential as serodiagnostic antigens using an enzyme-linked immunosorbent assay. When 100 sera from tuberculosis patients and 100 sera from the healthy controls were analyzed, rRV3369, rRv3874, and rRv0566c showed a sensitivity of 60{\%}, 74{\%}, and 43{\%}, and a specificity of 96{\%}, 97{\%}, and 84{\%}, respectively. These results suggest that the rRv3369 and rRv3874 proteins, which were expressed more abundantly in the more recently obtained clinical isolates of M. tuberculosis than in the laboratory-adapted H37Rv strain, are promising for use in the serodiagnosis of tuberculosis.",
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Antigens secreted from Mycobacterium tuberculosis : Identification by proteomics approach and test for diagnostic marker. / Young, Yil Bahk; Suk, Am Kim; Kim, Ji Soo; Euh, Hyung Jin; Bai, Gil Han; Cho, Sang Nae; Yu, Sam Kim.

In: Proteomics, Vol. 4, No. 11, 01.11.2004, p. 3299-3307.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Antigens secreted from Mycobacterium tuberculosis

T2 - Identification by proteomics approach and test for diagnostic marker

AU - Young, Yil Bahk

AU - Suk, Am Kim

AU - Kim, Ji Soo

AU - Euh, Hyung Jin

AU - Bai, Gil Han

AU - Cho, Sang Nae

AU - Yu, Sam Kim

PY - 2004/11/1

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AB - Tuberculosis caused by mycobacteria, mainly Mycobacterium tuberculosis, is a major infectious disease of the respiratory system. An early diagnosis followed by chemotherapy is the major control strategy. In an effort to identify the antigens suitable for immunodiagnosis and vaccines, the proteins secreted in a culture medium from the M. tuberculosis K-strain, which is the most prevalent among the clinical isolates in Korea and belongs to the Beijing family, were analyzed by two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) and compared with those from the M. tuberculosis H37Rv and CDC1551 strains. Eight proteins, Rv0652, Rv1636, Rv2818c, Rv3369, Rv3865, Rv0566c, MT3304, and Rv3160, were identified by matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS) or liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) and found to be relatively abundant in the culture medium from the M. tuberculosis K-strain but less so from the CDC1551 or H37Rv strains. In addition, Rv3874 (CFP-10), Rv-0560c and Rv3648c, which were expressed increasingly in the K and CDC1551 strains, were also identified using the same proteomics technology. All proteins were prepared by molecular cloning, expression in Escherichia coli followed by affinity purification. Among them, three proteins, rRv3369, rRv0566c, and rRv3874, were selected by prescreening and examined fortheir potential as serodiagnostic antigens using an enzyme-linked immunosorbent assay. When 100 sera from tuberculosis patients and 100 sera from the healthy controls were analyzed, rRV3369, rRv3874, and rRv0566c showed a sensitivity of 60%, 74%, and 43%, and a specificity of 96%, 97%, and 84%, respectively. These results suggest that the rRv3369 and rRv3874 proteins, which were expressed more abundantly in the more recently obtained clinical isolates of M. tuberculosis than in the laboratory-adapted H37Rv strain, are promising for use in the serodiagnosis of tuberculosis.

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