Low sensitivity of a urine LAM-ELISA in the diagnosis of pulmonary tuberculosis

<p>Abstract</p> <p>Background</p> <p>The development and evaluation of rapid and accurate new diagnostic tools is essential to improve tuberculosis (TB) control in developing countries. In a previous study, the first release of a urine LAM-ELISA by Chemogen (Portland, USA) has been evaluated with a promising sensitivity and specificity for the diagnosis of pulmonary TB. In the present study, the now commercially available assay has been clinically assessed regarding its diagnostic value alone and in combination with clinical co-factors.</p> <p>Methods</p> <p>The test was applied to two urine samples from 291 consecutively enrolled Tanzanian patients with suspected pulmonary tuberculosis. The participants were subsequently assigned to classification groups according to microbiological, clinical and radiological findings at recruitment and during a maximum follow up period of 56 days.</p> <p>Results</p> <p>Only 35 out of 69 pulmonary TB cases -confirmed by smear microscopy and/or solid culture and/or liquid culture- showed at least one positive LAM-ELISA result (sensitivity 50.7%). The sensitivity was noticeably higher in females (66.7%) and in HIV positive participants (62.0%). The specificity amounted to 87.8% and was determined in participants with negative results in all microbiological tests and with sustained recovery under antibiotic treatment at day 56. Correlation with urinalysis revealed that proteinuria was significantly and positively associated with LAM-positivity (<it>P </it>= 0.026).</p> <p>Conclusion</p> <p>This commercially available generation of LAM-ELISA does not appear to be useful as an independent diagnostic test for pulmonary tuberculosis. The question whether the assay is suitable as a supplemental device in the diagnosis of HIV-associated TB, requires further investigations.</p

Characterization of monoclonal antibodies to protein antigen of Salmonella typhi

Two monoclonal antibodies were produced against protein antigens of Salmonella typhi. One of the antibodies (STP14) belongs to the immunoglobulin G1K subclass, and the other (STP13) was assigned to the immunoglobulin G2a(kappa) subclass. Both antibodies could recognize the 34.0-kilodalton protein antigen from S. typhi. The specificity of these antibodies was tested by immunoblotting with a panel of crude protein antigens from 12 bacteria causing enteric fever and enteric fever-like illness: S. typhi, S. paratyphi A, S. paratyphi B, S. paratyphi C, S. choleraesuis, S. enteritidis, S. krefeld, S. panama, S. typhimurium, Escherichia coli, Pseudomonas pseudomallei, and Yersinia enterocolitica. In a modified double-antibody sandwich enzyme-linked immunosorbent assay they could detect the protein antigen at ca. 0.6 microgram/ml. These monoclonal antibodies should be of great value in the diagnostic test for detecting S. typhi antigen in samples of bodily fluids isolated from patients with typhoid fever and in studies of the chemical structure and other immunological properties of this 34.0-kilodalton protein.</jats:p

Monoclonal antibodies to 52-kilodalton protein of Salmonella typhi

Ten monoclonal antibodies (6 immunoglobulin G1 kappa [IgG1 kappa] and 4 IgG2b kappa) from six hybrid clones specific for Salmonella typhi antigen were produced by immunizing BALB/cJ mice with affinity-purified S. typhi proteins (Bp). The latter were prepared by passing crude S. typhi Bp through an affinity column made from Sepharose conjugated to IgG antibodies against partially purified S. typhi Bp. The eluent was subsequently used as the immunogen for the production of monoclonal antibody. All 10 monoclonal antibodies reacted specifically with a 52-kilodalton (kDa) protein of S. typhi and were species specific. The presence of IgM antibody to the 52-kDa antigen in the sera of a majority of patients with acute typhoid fever suggested that this 52-kDa protein is also a good immunogen for humans. The potential usefulness of this antigen in the early diagnosis of typhoid fever is discussed.</jats:p

Systemic and intestinal immunities after natural typhoid infection

A 2-year study of systemic and intestinal immunity to Salmonella typhi was performed in 14 patients who were suffering from typhoid fever in an attempt to extrapolate the mechanism of immune responses in this disease. The methods employed were the leukocyte migration inhibition agarose test for the measurement of systemic cell-mediated immunity. The systemic immunoglobulin G (IgG) and IgM that were specific to S. typhi and anti-O and anti-H agglutinins were measured by indirect enzyme-linked immunosorbent assay (ELISA) and the Widal test, respectively. The immunobead ELISA was used to measure total secretory IgA (SIgA), and indirect ELISA was used to measure SIgA that was specific to S. typhi in the intestinal lavage fluid. The results revealed that the patients developed various types of immune response to S. typhi that had various magnitudes and durations. After the onset of illness, the cell-mediated immunity persisted for 16 weeks; IgG, IgM, and anti-O and anti-H agglutinin persisted for 2 years, 16 weeks, 16 weeks, and 36 weeks, respectively. SIgA can persist in the gut for about 48 weeks. Thus, the immunities as a whole can barely persist beyond 1 year after the onset of illness, unless there are persistent booster stimulations by S. typhi bacilli that exist in the environment, and then the immunities may be lifelong.</jats:p

Serological approaches of Typhoid fever

[no abstract available