Serologic tests play an important role in diagnosis of
typhoid fever. In an effort to develop a more defined
reagent for these tests, purified Salmonella enterica serovar Typhi (ST) O:1,9,12
polysaccharide was conjugated to
human serum albumin (HSA), and the conjugate was purified chromatographically to yield a
reagent with 2 moles ST O
polysaccharide per mole HSA. In 40 patients with bacteriologically confirmed
typhoid fever, significant dot immunobinding titers (> or =20,000) were present in 28 (70%) tested with 100 ng of ST
O antigen-HSA (ST O-HSA) conjugate, in 38 (95%) tested with 100 ng of ST
lipopolysaccharide, and in 16 (40%) tested with purified unconjugated ST O chains. In sera from 22 patients with other nontyphoid
fevers, 2 (9.1%) had such reactivities with 100 ng of ST O-HSA, 1 (4.5%) had such reactivity with 100 ng of ST
lipopolysaccharide (4.5%), and none reacted with 100 ng of unconjugated ST O chains. None of the 17 healthy-control sera reacted significantly with any of the ST
reagents. None of the patient or control sera reacted with unconjugated HSA. The sensitivity of dot immunobinding for
typhoid fever was 70% with 100 ng of ST O-HSA, somewhat lower than that with 100 ng of ST
lipopolysaccharide (95%) but similar to that of the Widal H agglutination test with a > or =1/160 cutoff (74%). Specificities of these tests were 91%, 95%, and 86%, respectively. These preliminary results suggest that ST O
polysaccharide-
protein conjugates could provide a nontoxic, easily quality-controlled synthetic
reagent for analysis of human immune responses to ST as well as for the development of new diagnostics and
vaccines for
typhoid fever.