RESEARCH PAPER
Sequence based typing and pre-absorption test in retrospective analysis of a pseudo-outbreak of Legionella infections differentiates true cases of legionellosis
1
National Institute of Public Health-National Institute of Hygiene, Warsaw, Poland
2
Department of Immunology and Clinical Allergology, Military Institute of Medicine, Warsaw, Poland
3
Serum Production, SSI Diagnostica, Statens Serum Institute, Hillerod, Denmark
Ann Agric Environ Med. 2012;19(3):437-443
KEYWORDS
legionellosispseudo-outbreakfalse positive resultsLegionella serologypre-absorption testsequence base typing
ABSTRACT
The aim of this study was elimination of false positive results obtained by the Chlamylege kit. Two serological kits (IgM ELISA L.pneumophila sgs1-7; ImmuViewTM L.pneumophila sg1/sg3) and pre-absorption tests (with L.pneumophila sg1 and sg3 reference strains antigens) were used. 153 sera (79 patients) were examined. The high correlations were found between the results by both tests. Positive results by ELISA (sgs1-7) were found in 19/79 patients; by ImmuViewTM (sg1+sg3) in 16/63. In 8 patients, the dynamics of the IgM in pairs of sera was high (ratio ≥2). In 5/8 of those patients seroconversion was determined. Selected pre-absorbed sera (15 pairs) were tested simultaneously by the same tests. In 8/15 pairs of sera, the reduction of IgM levels in pre-absorbed sera was higher than 10. The reduction of IgM differed in sg1 and sg3 tests. The probability of infections due to L.pneumophila sg3 (7 patients) and L.pneumophila sg1 (5 patients) was based on the results of pre-absorption tests. The correlation between ELISA and ImmuViewTM tests of pre-absorbed sera was statistically significant (Po=0.0389). Moreover, genotyping of L.pneumophila (SBT) directly in the sera of selected 15 patients (high IgM reduction) was carried out. Completed 7 alleles profile (ST36) was determined in one patient. However, a second patient had the same profile of 5 alleles, and similar reactions in pre-absorption tests. At least 4 sources of infections were suggested on the base of genotyping and pre-absorption results. Conclusions:Positive results obtained by molecular techniques (eg.PCR) in the diagnosis of Legionella infections should be supplemented by other tests for confirmation of legionellosis. The sequence based typing carried out directly in clinical specimens seems to be a promising method.
REFERENCES (23)
2.
Modi A, Gardner J, Lighton L, Coetzee N. Pontiac fever outbreak associated with a spa-pool, United Kingdom, April 2008. Euro Surveill. 2008; 13 (30): pii=18934. http://www.eurosurveillance.or.... spx? ArticleId=18934.
3.
Pancer K, Napiórkowska A, Gut W, Stypułkowka-Misiurewicz H. Demographic characteristics of reported cases of legionellosis in years 2005-2009 in Poland in comparison to EWGLI data. Przegl Epidemiol. 2011; 65: 433-439.
4.
Joseph CA, Ricketts KD, on behalf of the European Working Group for Legionella Infections Legionnaires’ disease in Europe 2007–2008. Euro Surveill 2010; 15 (8): pii=19493 http://www.eurosurveillance.or... ViewArticle.aspx? ArticleId=19493.
5.
Joseph CA, Ricketts KD, Yadav R, Patel S, on behalf of the European Working Group for Legionella Infections Travel-associated Legionnaires’ disease in Europe in 2009. Euro Surveill 2010; 15 (41): pii=19683 http:// www.eurosurveillance.org/ViewArticle.aspx? ArticleId=19683.
6.
Licciardi PV, Balloch A, Russel FM, Mulholland EK, Tang MLK. Antibodies to serotype 9V exhibit novel serogroup cross-reactivity following infant pneumococcal immunization. Vaccine 2010; 28: 3793-3800.
7.
Brown-Augsburger P, Masnyk M, Santa P, Gingerich RL, Bowster RR. An immunoadsorption strategy to produce specific antisera against analogs of human proteins: development of sensitive and specific radioimmunoassays for two analogs of human leptin. J Pharm Biomed Anal. 2000; 23: 687-696.
8.
Alvarez J, Dominguez A, Sabria M, Ruiz L, Torner N, Cayla J et al. Impact of the Legionella urinary antigen test on epidemiological trends in community outbreaks of legionellosis in Catalonia, Spain, 1990—2004. Inter J Infect Dis. 2009; 13: e365–e370.
9.
Olsen CW, Elverdal P, Jørgensen CS, Uldum SA. Comparison of the sensitivity of the Legionella urinary antigen EIA kits from Binax and Biotest with urine from patients with infections caused by less common serogroups and subgroups of Legionella. Eur J Clin Microbiol Infect Dis. 2009; 28: 817–820.
10.
Javed S, Chaudhry R, Passi K, Sharma S, Padmaja K, Dhawan B et al. Sero diagnosis of Legionella infection in community acquired Pneumonia. Indian J Med Res. 2010; 131: 92-96.
11.
Elverdal PL, Svarrer CW, Jørgensen CS, Skovsted IC, Uldum SA. Development and validation of ELISA for detection of antibodies to Legionella pneumophila serogroup 1, 3 and 6 in human sera. J Microbiol Met. 2011; 86: 298–303.
12.
Rojas A, Navarro MD, Forne´s FE, Serra E, Simarro E, Rojas J et al. Value of Serological Testing for Diagnosis of Legionellosis in Outbreak Patients. J Clin Microbiol. 2005; 43: 4022-4025.
13.
Yzerman EPF, den Boer JW, Lettinga KD, Arnoud J. Schel AJ, Schellekens J et al. Sensitivity of three serum antibody tests in a large outbreak of Legionnaires’ disease in the Netherlands. J Med Microbiol. 2006; 55: 561–566.
14.
Kern M, Böhm S, Deml L, Wolf H, Reischl U, Niller HH. Inhibition of Legionella pneumophila PCR in respiratory samples: A quantitative approach. J Microbiol Met. 2009; 79: 189–193.
15.
Stojek NM, Dutkiewicz J. Co-existence of Legionella and other Gram-negative bacteria in potable water from various rural and urban sources. Ann Agric Environ Med 2011; 18: 330-334.
16.
von Baum H, Ewig S, Marre R, Suttorp N, Gonschior S, Welte T et al. Community-Acquired Legionella Pneumonia. New Insights from the German Competence Network for Community Acquired Pneumonia. Clin Infect Dis. 2008; 46: 1356–64.
17.
Carr R, Warren R, Towers L, Bartholomew A, Duggal HV, Rehman Y et al. Investigating a cluster of Legionnaires’cases: Public health implications. Public Health. 2010; 124: 326-331.
18.
Borchardt J, Helbig JH, Lück PC. Occurrence and distribution of sequence types among Legionella pneumophila strains isolated from patients in Germany common features and differences to other regions of the world. Eur J Clin Microbiol Infect Dis. 2008; 27: 29–36.
19.
Chasqueira MJ, Rodrigues L, Nascimento M, Marques T. Sequence-based and monoclonal antibody typing of Legionella pneumophila isolated from patients in Portugal during 1987-2008. Euro Surveill. 2009; 14 (28): pii=19271. http://www.eurosurveillance.or.... aspx? ArticleId=19271.
20.
Harrison TG, Afshar B, Doshi N, Fry NK, Lee JV. Distribution of Legionella pneumophila serogroups, monoclonal antibody subgroups and DNA sequence types in recent clinical and environmental isolates from England and Wales (2000–2008). Eur J Clin Microbiol Infect Dis. 2009; 28: 781–791.
21.
Pravinkumar SJ, Edwards G, Lindsay D, Redmond S, Stirling J, House R et al. A cluster of Legionnaires’ disease caused by Legionella longbeachae linked to potting compost in Scotland, 2008-2009. Euro Surveill. 2010; 15 (8): pii=19496. http://www.eurosurveillance.or... ViewArticle.aspx? ArticleId=19496.
22.
Reimer AR, Au S, Schindle S, Bernard KA. Legionella pneumophila monoclonal antibody subgroups and DNA sequence types isolated in Canada between 1981 and 2009: Laboratory Component of National Surveillance. Eur J Clin Microbiol Infect Dis. 2010; 29: 191–205.
23.
Campese C, Bitar D, Jarraud S, Maine C, Forey F, Etienne J et al. Progress in the surveillance and control of Legionella infection in France, 1998–2008. Inter J Infect Dis. 2011; 15: e30–e37.
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