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(1) Background: Tuberculosis (TB) in humans is a serious chronic epidemic disease caused by Mycobacterium tuberculosis ( M. tb ). The diagnosis of TB, especially extra-pulmonary TB (EPTB), is difficult. Isolation of M. tb from culture has a low sensitivity in patients with TB and an even lower sensitivity in cases of EPTB. Although Xpert MTB/RIF assays and serological tests are more sensitive than the above tests, they still lack sensitivity for EPTB diagnosis. (2) Methods: To improve the accuracy of TB diagnosis, a Rv0222-Rv2657c-Rv1509 fusion protein based iELISA was constructed, the diagnosis of TB, pulmonary TB (PTB) and EPTB was then evaluated. Sera of 40 TB patients including 14 with PTB, 14 with EPTB and 12 with no information about the form of TB, and five pneumonia patients were investigated. (3) Results: The sensitivity of the ELISA in TB, PTB and EPTB patients was 80% (95% CI: 64.4, 90.9%), 85.7% (95% CI: 57.2, 98.2%) and 92.8% (95% CI: 66.1, 99.8%), respectively, with a specificity of 70% (95% CI: 53.5, 83.4%). Both the sensitivity and specificity with this fusion protein were higher than for CFP10/ESAT6 (used as reference antigen) fusion protein (71.4%; 95% CI: 41.9, 91.6%, and 67.5%; 95% CI: 50.9, 81.4%), respectively, in cases of EPTB. All pneumonia patients' sera tested negative in both ELISAs. (4) Conclusion: use of these new fusion proteins as antigens in serological assays has the potential to improve the diagnosis of all forms of TB in humans, especially EPTB.

Use of Rv0222-Rv2657c-Rv1509 Fusion Protein to Improve the Accuracy of an Antibody ELISA for Extra-Pulmonary Tuberculosis in Humans