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Journal of Clinical Microbiology, September 2005, p. 4545-4550, Vol. 43, No. 9
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.9.4545-4550.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Salmonella enterica Serovar Typhi O:1,9,12 Polysaccharide-Protein Conjugate as a Diagnostic Tool for Typhoid Fever

Jessica Zuñiga,1 Luis Lillo,1 Junghee J. Shin,2 Rajya L. Machavarapu,2 Teresa Quiroga,3 Manuela Goycoolea,3 Betty Matsuhiro,1 L. Aron-Hott,2 Henry P. Godfrey,4* and Felipe C. Cabello2

Department of Chemistry, University of Santiago de Chile, Santiago, Chile,1 Department of Microbiology and Immunology, New York Medical College, Valhalla, New York 10595,2 Central Laboratory, Clinical Laboratories Diagnostic Center, School of Medicine, Pontificia Catholic University of Chile, Santiago, Chile,3 Department of Pathology, New York Medical College, Valhalla, New York 105954

Received 27 July 2004/ Returned for modification 2 December 2005/ Accepted 1 June 2005

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 (≥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 ≥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.

* Corresponding author. Mailing address: Department of Pathology, Basic Science Building, New York Medical College, Valhalla, NY 10595-1690. Phone: (914) 594-4160. Fax: (914) 594-4163. E-mail: hgodfrey{at}nymc.edu.

Journal of Clinical Microbiology, September 2005, p. 4545-4550, Vol. 43, No. 9
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.9.4545-4550.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.





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