Serological detection of Mycobacterium Tuberculosis complex infection in multiple hosts by One Universal ELISA

Abstract

Tuberculosis (TB), a contagious disease mainly caused by Mycobacterium tuberculosis (M. tb), Mycobacterium bovis (M. bovis), and Mycobacterium caprae (M. caprae), poses a major global threat to the health of humans and many species of animals. Developing an ante-mortem detection technique for different species would be of significance in improving the surveillance employing a One Health strategy. To achieve this goal, a universal indirect ELISA was established for serologically detecting Mycobacterium tuberculosis complex infection for multiple live hosts by using a fusion protein of MPB70, MPB83, ESAT6, and CFP10 common in M. tb, M. bovis, and M. caprae as the coating antigen (MMEC) and HRP-labeled fusion protein A and G as a secondary antibody. After testing the known positive and negative sera, the receiver operating characteristic curves were constructed to decide the cut-off values. Then, the diagnostic sensitivity and specificity of MMEC/AG-iELISA were determined as 100.00% (95% CI: 96.90%, 100.00%) and 100.00% (95% CI: 98.44%, 100.00%) for M. bovis infection of cattle, 100.00% (95% CI: 95.00%, 100.00%) and 100.0% (95% CI: 96.80%, 100.00%) for M. bovis infection of sheep, 90.74% (95% CI: 80.09%, 95.98%) and 98.63% (95% CI: 95.14%, 99.76%) for M. bovis infection of cervids, 100.00% (95% CI: 15.81%, 100.00%) and 98.81% (95% CI: 93.54%, 99.97%) for M. bovis infection of monkeys, 100.00% (95% CI: 86.82%, 100.00%) and 94.85% (95% CI: 91.22%, 97.03%) for M. tb infection of humans. Furthermore, this MMEC/AG-iELISA likely detects M. caprae infection in roe deer. Thus this method has a promising application in serological TB surveillance for multiple animal species thereby providing evidence for taking further action in TB control.

Comparative diagnosis of bovine tuberculosis using single intradermal cervical tuberculin technique, conventional methods, enzyme-linked immunosorbent assay, and the gamma-interferon assay

“…Antibodies are only produced during the advanced phases of TB and usually cannot be detected in the early stages of the disease [ 31 ]. Single antigen testing also provides low detection sensitivity [ 6 ]. However, ELISA can be used to monitor the progress of infection [ 32 ] and can be used to confirm the results of skin tests [ 33 ].…”

“…The tuberculin test is the test of choice globally and the most common test for bovine TB and international trade [ 5 ]. However, the tuberculin test is limited by variable specificity and sensitivity due to common skin reactive antigens that cause reactions in animals sensitized to non-tuberculous mycobacteria [ 6 ]. Others disadvantages of tuberculin test include the individual variation in test performance in injection, measurement ability, and test interpretation [ 7 ].…”

Evolution of tuberculosis diagnostics: From molecular strategies to nanodiagnostics

“…Infection can be detected by using fusion protein MMEC (MPB70, MPB83, ESAT6, and CFP10 i.e., common in different species of mycobacteria) as coating antigen. The sensitivity and specificity of this ELISA is 100% and 94.85% respectively for Mtb detection in humans, making it a reliable procedure for TB detection [ 65 ].…”

Spotlight on mycobacterial lipid exploitation using nanotechnology for diagnosis, vaccines, and treatments

“… 4. , 5. It is noteworthy that the different lineages of these species, which are part of the M. tuberculosis complex (MTC), 4 display phenotypic differences that should be considered in the design of control measures.…”