Improved detection of mycobacterial antigens in clinical specimens by combined enzyme-linked immunosorbent assay

Abstract

Three types of antibodies against cellular and secretory-excretory protein antigens were simultaneously used for the direct detection of mycobacterial antigens in sputum and cerebrospinal fluid (CSF) specimens, using enzyme-linked immunosorbent assay (ELISA). The antibodies consisted of in-house raised and prepared anti-whole-cell, heat-killed, and sonicated Mycobacterium tuberculosis, anti-secretory-excretory protein extract of bacilli Calmette-Guerin (BCG) strain, and commercially available anti-BCG. Sputum specimens comprised 24 smear positive, culture positive, and 47 smear-negative, culture positive (SNCP), from patients with pulmonary tuberculosis, as well as 45 smear-negative, culture-negative (SNCN) control samples. The CSF specimens included 18 SNCPs from patients with tuberculous meningitis and 18 SNCN controls. The sensitivity of the individual tests for sputum and CSF specimens ranged from 70% to 79% and 72% to 89%, respectively, whereas in the combined tests it reached 86%-96% for sputum specimens and 100% for CSF specimens. The specificity of ELISAs for sputum specimens was lower in the combined (73%-87%) than in the individual (87%-98%) tests, whereas for CSF specimens it was 100% in all tests. Thus, the combined ELISA approach for mycobacterial antigen detection provides a rapid and reliable laboratory adjunct in the diagnosis of patients with tuberculosis.