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Clinical and Vaccine Immunology, December 2008, p. 1788-1795, Vol. 15, No. 12
1071-412X/08/$08.00+0     doi:10.1128/CVI.00231-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Improved Sensitivity of Diagnosis of Tuberculosis in Patients in Korea via a Cocktail Enzyme-Linked Immunosorbent Assay Containing the Abundantly Expressed Antigens of the K Strain of Mycobacterium tuberculosis{triangledown}

A-Rum Shin,1 Sung Jae Shin,1* Kil-Soo Lee,1 Sun-Ho Eom,1 Seung-Sub Lee,1 Byung-Soo Lee,1 Ji-Sook Lee,1 Sang Nae Cho,2 and Hwa-Jung Kim1*

Department of Microbiology and Infection Signaling Network Research Center, College of Medicine, Chungnam National University, Daejeon,1 Department of Microbiology and Institute of Immunology and Immunological Diseases, Yonsei University College of Medicine, Seoul, South Korea2

Received 21 June 2008/ Returned for modification 23 July 2008/ Accepted 28 September 2008

Tuberculosis (TB) is the leading cause of death from a single infectious agent in Korea. In this study, we compared the proteins present in culture filtrates from Mycobacterium tuberculosis strain K, which is the dominant clinical isolate in Korea, with those present in culture filtrates from M. tuberculosis H37Rv. Several differences in expression were detected between the two strains for those proteins with a molecular mass of <20 kDa. ESAT-6, HSP-X, and CFP-10 were found to be abundantly expressed in the strain K culture filtrates by liquid chromatography-electrospray ionization-time of flight mass spectrometry. The serodiagnostic potentials of recombinant antigens rESAT-6, rHSP-X, and rCFP-10 and two native antigens (Ag85 and PstS1) were evaluated by Western blot analysis and enzyme-linked immunosorbent assay (ELISA) using sera collected from 46 TB patients with active disease and 46 healthy controls. As for our ELISA results, HSP-X was superior to the other antigens in terms of sensitivity when a single antigen was employed. The results of a receiver operator characteristic analysis revealed that a cocktail ELISA using all five antigens was significantly more sensitive (77.8%) than the use of a single antigen and offered equivalent specificity; moreover, it produced the largest area under the curve (0.91 versus 0.55 to 0.87). Therefore, a cocktail ELISA containing abundantly expressed antigens enhances the sensitivity of a single antigen and can be a useful diagnostic tool for the detection of active TB.

* Corresponding author. Mailing address: Department of Microbiology, College of Medicine, Chungnam National University, 6 Munhwa-dong, Jungku, Daejeon 301-747, South Korea. Phone for Hwa-Jung Kim: 82 42 580 8242. Fax: 82 42 585 3686. E-mail: hjukim{at}cnu.ac.kr. Phone for Sung Jae Shin: 82 42 580 8246. E-mail: sjshin{at}cnu.ac.kr

{triangledown} Published ahead of print on 22 October 2008.

Clinical and Vaccine Immunology, December 2008, p. 1788-1795, Vol. 15, No. 12
1071-412X/08/$08.00+0     doi:10.1128/CVI.00231-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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