Immunological evaluation of a 12-kilodalton protein of Mycobacterium tuberculosis by enzyme-linked immunosorbent assay

ABSTRACT 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.

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False-positive reactions with enzyme-linked immunosorbent assay of Mycobacterium tuberculosis antigens in pleural fluid

Journal of Medical Microbiology ◽

10.1099/00222615-26-4-241 ◽

1988 ◽

Vol 26 (4) ◽

pp. 241-243 ◽

Cited By ~ 14

Author(s):

R. Dhand ◽

N. K. Ganguly ◽

C. Vaishnavi ◽

R. Gilhotra ◽

S. K. Malik

Keyword(s):

Mycobacterium Tuberculosis ◽

Pleural Fluid ◽

Immunosorbent Assay ◽

False Positive ◽

Enzyme Linked Immunosorbent Assay ◽

False Positive Reactions

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Heterogeneous Antibody Responses in Tuberculosis

Infection and Immunity ◽

10.1128/iai.66.8.3936-3940.1998 ◽

1998 ◽

Vol 66 (8) ◽

pp. 3936-3940 ◽

Cited By ~ 162

Author(s):

Konstantin Lyashchenko ◽

Roberto Colangeli ◽

Michel Houde ◽

Hamdan Al Jahdali ◽

Dick Menzies ◽

...

Keyword(s):

Mycobacterium Tuberculosis ◽

Antibody Response ◽

Immunosorbent Assay ◽

Immunoglobulin G ◽

Antigen Recognition ◽

Antibody Responses ◽

Enzyme Linked Immunosorbent Assay ◽

Protein Antigens ◽

Tuberculosis Patients ◽

Specific Antibodies

ABSTRACT Antibody responses during tuberculosis were analyzed by an enzyme-linked immunosorbent assay with a panel of 10 protein antigens of Mycobacterium tuberculosis. It was shown that serum immunoglobulin G antibodies were produced against a variety of M. tuberculosis antigens and that the vast majority of sera from tuberculosis patients contained antibodies against one or more M. tuberculosis antigens. The number and the species of serologically reactive antigens varied greatly from individual to individual. In a given serum, the level of specific antibodies also varied with the antigen irrespective of the total number of antigens recognized by that particular serum. These findings indicate that person-to-person heterogeneity of antigen recognition, rather than recognition of particular antigens, is a key attribute of the antibody response in tuberculosis.

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Genotyping ofSalmonellaspp. on the basis of CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)

10.1101/473868 ◽

2018 ◽

Author(s):

Calvin Jiksing ◽

Normah Yusop ◽

Farhan Nazaie Nasib ◽

Kenneth Francis Rodrigues

Keyword(s):

Polymerase Chain Reaction ◽

Mycobacterium Tuberculosis ◽

Immunosorbent Assay ◽

Deoxyribonucleic Acid ◽

Enzyme Linked Immunosorbent Assay ◽

Chain Reaction ◽

Pcr Polymerase Chain Reaction ◽

Crispr Array ◽

Polymerase Chain ◽

Veterinary Laboratory

ABSTRACTAimsBacterial genotyping on the basis of the CRISPR array has been established inMycobacterium tuberculosiswith a method called spacer oligonucleotide typing (spoligotyping). The spoligotyping method had been widely used for both detection and typing ofM. tuberculosiscomplex bacteria. This present study aimed at determining if the CRISPR array inSalmonellaspp. could be applied to establish a correlationship between serogroup and the fingerprint generated by CRISPR typing.Methodology and resultsA total of 30 samples were obtained from Diagnostic Veterinary Laboratory, Kota Kinabalu, Sabah. Serogroup was determined on the basis of ELISA (enzyme-linked immunosorbent assay). Four different serogroups were identified which were serogroup B, C, D, and E. DNA (deoxyribonucleic acid) was extracted and PCR (polymerase chain reaction) was performed using primers which were designed to amplify the CRISPR array inSalmonellagenome. Our results indicate that there is a correlationship between serogroup obtained using ELISA and the profile generated by CRISPR typing.Conclusion, significance and impact of studyCRISPR typing has the potential to be applied for the genotyping ofSalmonella.

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