Rapid decay of Salmonella flagella antibodies during human gastroenteritis: A follow up study

https://doi.org/10.1016/j.mimet.2005.02.006Get rights and content

Abstract

An indirect enzyme-linked immunosorbent assay (ELISA) based on Salmonella re-polymerized flagella was employed to measure levels of immunoglobulin (Ig) G, IgM and IgA antibodies in sera from 303 Danish patients diagnosed with either Salmonella enteritidis or Salmonella typhimurium. The antibody-levels were assessed at one, three and six months after onset of salmonellosis, and sera from a control-group of 170 healthy blood donors were additionally analysed in order to establish cut-off values for the analysis. Cross-reactions to other Salmonella serotypes, as well as to Escherichia coli, Yersinia enterocolitica, Campylobacter jejuni, Campylobacter coli and Helicobacter pylori were observed. At one month after onset of symptoms, 70% of the patients recovering from a S. enteritidis infection carried detectable levels of anti-flagella antibodies, as did 77% of the patients recovering from S. typhimurium infection. Three months after onset of symptoms these detection rates had decreased to 46% and 40%; and six months after onset of symptoms the detection rates were 34% and 38%. This rapid decrease in the serum levels of flagella antibodies is in conflict with the “common knowledge” statement of a long-lasting anti-flagella immunoresponse. The present study suggests that such a tenacious statement is (or may be) inaccurate.

Introduction

In Denmark, infections by Salmonella enteritidis or Salmonella typhimurium are the second and third most common cause of bacterial gastrointestinal infections, with Campylobacter being the most common cause (Anonymous, 2003).

At such gastrointestinal infections with Salmonella, the symptoms are diarrhea, vomiting, fever and abdominal pain; all of which commonly occur within 48 h after ingestion of the bacteria. The most common vectors of infection are meat, dairy products, poultry and eggs (Anonymous, 2003, Hohmann, 2001, Santos et al., 2001). At suspicion of infection with Salmonella, diagnosis is done by fecal or blood-culturing and/or detection of specific antibodies.

Flagella are hair like structures scattered on the surface of the Salmonella cell and the use of flagella-antigens in agglutination assays or ELISAs for detection of human typhoid fever (Shukla et al., 1997, Parry et al., 1999, Jesudason et al., 1998, Frimpong et al., 2000) and non-typhoid salmonellosis in chickens and livestock (Timoney et al., 1990, Gast and Holt, 2001, Yap et al., 2001, Feberwee et al., 2001, Veling et al., 2001) is a frequently used procedure. However, the present study is the first to employ flagella-antigens in an ELISA to investigate the magnitude and timeline of the human antibody-response against Salmonella-flagella at non-typhoid salmonellosis.

The antibody-response against Salmonella flagella is generally believed to be long-lasting (Parry et al., 1999, Frimpong et al., 2000, Willke et al., 2002, Schroeder, 1968), and a flagella-based ELISA might therefore be a means of post-infection diagnostics. But, as presented below, this hypothesis is questionable.

Section snippets

Patients

Sera from 303 Danish patients with gastroenteritis diagnosed by fecal culturing with infection by either S. enteritidis (153 patients) or S. typhimurium (150 patients) were available from a previous study. According to the laboratory-based national surveillance data for gastrointestinal infections, none of these patients were notified as having suffered from gastroenteritis within the previous year. All patients were asked to deliver a blood-sample shortly after the time of diagnosis, as well

Quality of antigens

SDS-PAGE analyses of the purified S. godesberg and S. mgulani antigens revealed two distinct fragments for both as well as a few vague fragments (Fig. 1). This has also been observed by others (Iankov et al., 2002, Chart et al., 2002, Timoney et al., 1990, Zamora et al., 1999, Ibrahim et al., 1985, Chart et al., 1997, Sbrogio-Almeida and Ferreira, 2001). For the S. godesberg flagellin, the two distinct fragments as well as one of the vague fragments were analysed by mass spectrometry,

Discussion

Patients suffering from infections with either S. enteritidis or S. typhimurium were found to produce detectable antibodies against the Salmonella flagella when their sera were analysed by ELISA. At approximately one month after onset of salmonellosis symptoms, 70% of the S. enteritidis patients were found positive for the presence of H:[g,m] antibodies, and 77% of the S. typhimurium were likewise found positive for the presence of H:[i:1,2] antibodies—in comparison, the sensitivities obtained

Acknowledgements

This study was partially supported by the Ministry of Food Agriculture and Fisheries, Directorate for Food Fisheries and Agro Business (FØS100-SSI-7).

References (29)

  • E.H. Frimpong et al.

    Determination of diagnostic Widal titres in Kumasi, Ghana

    West Afr. J. Med.

    (2000)
  • R.K. Gast et al.

    The relationship between the magnitude of the specific antibody response to experimental Salmonella enteritidis infection in laying hens and their production of contaminated eggs

    Avian Dis.

    (2001)
  • J. Gobom et al.

    Sample purification and preparation technique based on nano-scale reversed-phase columns for the sensitive analysis of complex peptide mixtures by matrix-assisted laser desorption/ionization mass spectrometry

    J. Mass Spectrom.

    (1999)
  • E.L. Hohmann

    Nontyphoidal salmonellosis

    Clin. Infect. Dis.

    (2001)
  • Cited by (14)

    • Production of recombinant flagellin to develop ELISA-based detection of Salmonella Enteritidis

      2017, Brazilian Journal of Microbiology
      Citation Excerpt :

      As such, several screening methods such as nucleic acid-based assays and immunology-based assays have been examined for a rapid detection of Salmonella in the recent decades.20,22–24 In earlier studies, lipopolysaccharides (LPS) present on the bacterial surface were used in the ELISA-assay for the detection of Salmonella, but the problem with lipopolysaccharides is the cross-reactivity with other gram-negative bacteria.25–27 Therefore, flagellin (FliC) was sought as an alternative for rapid diagnosis of Salmonella and, during the last decade, several studies have used it for the detection of Salmonella.

    • Serological follow-up after non-typhoid salmonella infection in humans using a mixed lipopolysaccharide ELISA

      2013, International Journal of Medical Microbiology
      Citation Excerpt :

      The mixed ELISA had a high sensitivity of 92% for the detection of S. Enteritidis infection and 86% for S. Typhimurium infection in the first 30 days after symptom onset (Table 2). Diagnostic accuracy is by far superior to the Widal agglutination assay (Strid et al., 2007) and to a flagella antigen based ELISA previously evaluated by our group (Dalby et al., 2005). IgA and IgM were the most sensitive diagnostic parameters in the first 30 days.

    • Inferring the time of infection from serological data

      2019, Handbook of Infectious Disease Data Analysis
    View all citing articles on Scopus
    View full text