Serodiagnosis of Salmonella enterica serovar Typhi and S. enterica serovars Paratyphi A, B and C human infections

Author:

Chart Henrik1,Cheasty Thomas1,de Pinna Elizabeth1,Siorvanes Lisa1,Wain John2,Alam Didar3,Nizami Qamarauddin3,Bhutta Zulfiqar3,Threlfall E. John1

Affiliation:

1. Laboratory of Enteric Pathogens, Department of Gastrointestinal Infections, Centre for Infections, Health Protection Agency, 61 Colindale Avenue, London NW9 5EQ, UK

2. The Wellcome Trust Sanger Institute Wellcome Trust Genome Campus, Hinxton, Cambridge CB10 1SA, UK

3. Department of Paediatrics and Child Health, The Aga Khan University Medical Centre, PO Box 3500, Karachi, Pakistan

Abstract

The aim of this study was to evaluate an immunoassay for the detection of human serum antibodies to the LPS and flagellar antigens of Salmonella Typhi and Salmonella Paratyphi A, B and C, and to the Vi capsular polysaccharide of S. Typhi and S. Paratyphi C. A total of 330 sera were used; these originated from 15 patients who were culture-positive for S. Typhi and 15 healthy controls, together with 300 sera submitted to the Laboratory of Enteric Pathogens for Salmonella serodiagnosis. By SDS-PAGE/immunoblotting, all 15 sera from culture-positive patients had serum antibodies to the 9,12 LPS antigens and 10 had antibodies to the ‘d’ flagellar antigens. Of the 300 reference sera, 22 had antibodies to the 9,12 LPS antigens, one to the 1,4,5,12 LPS antigens and 12 to the 6,7 LPS antigens. Only two sera had antibodies to flagellar antigens, one of which bound to the ‘b’ and the other to the ‘d’ antigen. An ELISA was developed that successfully detected serum antibodies to the Vi capsular polysaccharides, but because of the kinetics of serum antibody production to the Vi, these antibodies may be of limited value in the serodiagnosis of acute infection with S. Typhi and S. Paratyphi C. The immunoassays described here provide a sensitive means of detecting serum antibodies to the LPS, flagellar and Vi antigens of S. Typhi and S. Paratyphi, and constitute a viable replacement for the Widal assay for the screening of sera. The Salmonella serodiagnosis protocols described here are the new standard operating procedures used by the Health Protection Agency's National Salmonella Reference Centre based in the Laboratory of Enteric Pathogens, Colindale, UK.

Publisher

Microbiology Society

Subject

Microbiology (medical),General Medicine,Microbiology

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