Enhanced serodiagnostic potential of a fusion molecule consisting of Rv1793, Rv2628 and a truncated Rv2608 of Mycobacterium tuberculosis

• Published in: PloS one 2021-11, Vol.16 (11), p.e0258389-e0258389
• Main Authors: Sulman, Sadaf, Shahid, Saher, Khaliq, Aasia, Ambreen, Atiqa, Khan, Imran H, Cooper, Andrea M, Akhtar, Muhammad Waheed
• Format: Article
• Language: English
• Subjects: Adolescent; Adult; Aged; Aged, 80 and over; Analysis; Antibodies; Antibodies, Bacterial - blood; Antigenic determinants; Antigens; Antigens, Bacterial - immunology; Bacterial Proteins - genetics; Bacterial Proteins - immunology; BCG; BCG vaccines; Biology and Life Sciences; Biomarkers; Care and treatment; Cloning; Diagnosis; Dosage and administration; E coli; Epitopes; Epitopes - immunology; Escherichia coli - genetics; Escherichia coli - metabolism; Female; Follow-Up Studies; Fusion protein; Humans; Male; Medical diagnosis; Medicine and Health Sciences; Middle Aged; Mycobacterium tuberculosis; Mycobacterium tuberculosis - immunology; Mycobacterium tuberculosis - isolation & purification; Pakistan - epidemiology; Plasmids; Proteins; Recombinant Fusion Proteins - immunology; Research and Analysis Methods; Risk factors; Sensitivity; Sensitivity and Specificity; Serologic Tests - methods; Serology; Sex; Sputum; Sputum - microbiology; Tuberculosis; Tuberculosis, Pulmonary - blood; Tuberculosis, Pulmonary - diagnosis; Tuberculosis, Pulmonary - epidemiology; Tuberculosis, Pulmonary - microbiology; Viral antibodies; Young Adult
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0258389

Serodiagnosis of tuberculosis (TB) can be rapid, reliable and cost-effective if the issue of variable antibody responses of TB patients against different Mycobacterium tuberculosis (Mtb) antigens can be overcome by developing fusion proteins containing epitopes from multiple antigens of Mtb. In this study, Mtb antigens Rv1793, Rv2628, Rv2608 and a truncated variant produced by removing non-epitopic region from N-terminal of Rv2608 (tnRv2608), and the fusion protein Rv1793-Rv2628-tnRv2608 (TriFu64), were expressed in E. coli and purified. Plasma samples from TB patients characterized by sex, age and sputum/culture positivity, were used to compare the sensitivity of the single antigens with the fusion protein. Sensitivity of Rv1793, Rv2628 and Rv2608, was 27.8%, 39% and 36.3%, respectively. Truncation of Rv2608 increased sensitivity by approximately 35% in confirmed TB cases. Sensitivity of the fusion construct, TriFu64 increased to 66% with a specificity of 100%. Importantly, tnRv2608 was better able to detect sputum and culture negative patients, and this carried through to the fusion protein. We demonstrate that fusion of Mtb proteins ensures broad sensitivity across disease types, sex and age groups in a Pakistani population.