Skip to main content
Top
Published in: Infection 6/2014

01-12-2014 | Clinical and Epidemiological Study

Gastrointestinal pathogens detected by multiplex nucleic acid amplification testing in stools of pediatric patients and patients returning from the tropics

Authors: C. Beckmann, U. Heininger, H. Marti, H. H. Hirsch

Published in: Infection | Issue 6/2014

Login to get access

Abstract

Background

Gastrointestinal infections are caused by a broad spectrum of pathogens. Conventional diagnostic procedures are resource and time consuming due to single pathogen testing, often in different laboratories.

Method

We analyzed 312 consecutive stool samples from pediatric patients (n = 127) with gastroenteritis or from adult travelers returning from the tropics with suspected parasite infestation (n = 185) using commercial multiplex nucleic acid amplification testing (NAT) (xTAG gastrointestinal pathogen panel, Luminex) covering 15 diarrhea-causing pathogens. The results of the positive samples and a representative number of negative samples were compared to standard methods, including NAT, direct antigen detection (DAD), bacterial culture and microscopy.

Results

Of the 185 samples from adult travelers, 21 (11 %) were multiplexNAT-positive, with enterotoxigenic Escherichia coli (4 %) being the predominant pathogen. Microscopic examination revealed Blastocystis hominis in 23 % not covered by the panel. MultiplexNAT scored positive in 66 pediatric samples (52 %), with rotavirus (27 %) being the most prevalent. All adenovirus-, rotavirus-, Clostridium difficile- and Cryptosporidium-positive samples were confirmed in external laboratories, but only 40 % of norovirus- and 29 % of Giardia-positive samples. Analysis of frozen specimens by bacterial culture showed the highest discrepancies with the multiplexNAT.

Conclusion

Our study demonstrates broad detection of relevant gastroenteritis pathogens by multiplexNAT with a short turnaround time. This is important for diagnosis, infection control and empiric management of gastroenteritis patients, but may be selectively complemented by bacterial culture and resistance testing.
Literature
1.
go back to reference Barbut F, Corthier G, Charpak Y, et al. Prevalence and pathogenicity of Clostridium difficile in hospitalized patients. A French multicenter study. Arch Intern Med. 1996;156:1449–54.PubMedCrossRef Barbut F, Corthier G, Charpak Y, et al. Prevalence and pathogenicity of Clostridium difficile in hospitalized patients. A French multicenter study. Arch Intern Med. 1996;156:1449–54.PubMedCrossRef
2.
go back to reference Barnes GL, Uren E, Stevens KB, et al. Etiology of acute gastroenteritis in hospitalized children in Melbourne, Australia, from April 1980 to March 1993. J Clin Microbiol. 1998;36:133–8.PubMedCentralPubMed Barnes GL, Uren E, Stevens KB, et al. Etiology of acute gastroenteritis in hospitalized children in Melbourne, Australia, from April 1980 to March 1993. J Clin Microbiol. 1998;36:133–8.PubMedCentralPubMed
3.
go back to reference Bernier C, Gounon P, Le Bouguenec C. Identification of an aggregative adhesion fimbria (AAF) type III-encoding operon in enteroaggregative Escherichia coli as a sensitive probe for detecting the AAF-encoding operon family. Infect Immun. 2002;70:4302–11.PubMedCentralPubMedCrossRef Bernier C, Gounon P, Le Bouguenec C. Identification of an aggregative adhesion fimbria (AAF) type III-encoding operon in enteroaggregative Escherichia coli as a sensitive probe for detecting the AAF-encoding operon family. Infect Immun. 2002;70:4302–11.PubMedCentralPubMedCrossRef
4.
go back to reference Black RE. Epidemiology of travelers’ diarrhea and relative importance of various pathogens. Rev Infect Dis. 1990;12:S73–9.PubMedCrossRef Black RE. Epidemiology of travelers’ diarrhea and relative importance of various pathogens. Rev Infect Dis. 1990;12:S73–9.PubMedCrossRef
5.
go back to reference Boisen N, Scheutz F, Rasko DA, et al. Genomic characterization of enteroaggregative Escherichia coli from children in Mali. J Infect Dis. 2012;205:431–44.PubMedCentralPubMedCrossRef Boisen N, Scheutz F, Rasko DA, et al. Genomic characterization of enteroaggregative Escherichia coli from children in Mali. J Infect Dis. 2012;205:431–44.PubMedCentralPubMedCrossRef
6.
go back to reference Bon F, Fascia P, Dauvergne M, et al. Prevalence of group A rotavirus, human calicivirus, astrovirus, and adenovirus type 40 and 41 infections among children with acute gastroenteritis in Dijon, France. J Clin Microbiol. 1999;37:3055–8.PubMedCentralPubMed Bon F, Fascia P, Dauvergne M, et al. Prevalence of group A rotavirus, human calicivirus, astrovirus, and adenovirus type 40 and 41 infections among children with acute gastroenteritis in Dijon, France. J Clin Microbiol. 1999;37:3055–8.PubMedCentralPubMed
7.
go back to reference Calderaro A, Gorrini C, Montecchini S, et al. Evaluation of a real-time polymerase chain reaction assay for the laboratory diagnosis of giardiasis. Diagn Microbiol Infect Dis. 2010;66:261–7.PubMedCrossRef Calderaro A, Gorrini C, Montecchini S, et al. Evaluation of a real-time polymerase chain reaction assay for the laboratory diagnosis of giardiasis. Diagn Microbiol Infect Dis. 2010;66:261–7.PubMedCrossRef
9.
go back to reference Cohen SH, Gerding DN, Johnson S, et al. Clinical practice guidelines for Clostridium difficile infection in adults: 2010 update by the society for healthcare epidemiology of America (SHEA) and the infectious diseases society of America (IDSA). Infect Control Hosp Epidemiol. 2010;31:431–55.PubMedCrossRef Cohen SH, Gerding DN, Johnson S, et al. Clinical practice guidelines for Clostridium difficile infection in adults: 2010 update by the society for healthcare epidemiology of America (SHEA) and the infectious diseases society of America (IDSA). Infect Control Hosp Epidemiol. 2010;31:431–55.PubMedCrossRef
10.
go back to reference Coste JF, Vuiblet V, Moustapha B, et al. Microbiological diagnosis of severe diarrhea in kidney transplant recipients by use of multiplex PCR assays. J Clin Microbiol. 2013;51:1841–9.PubMedCentralPubMedCrossRef Coste JF, Vuiblet V, Moustapha B, et al. Microbiological diagnosis of severe diarrhea in kidney transplant recipients by use of multiplex PCR assays. J Clin Microbiol. 2013;51:1841–9.PubMedCentralPubMedCrossRef
11.
12.
13.
go back to reference Farkas T, Jiang X, Guerrero ML, et al. Prevalence and genetic diversity of human caliciviruses (HuCVs) in Mexican children. J Med Virol. 2000;62:217–23.PubMedCrossRef Farkas T, Jiang X, Guerrero ML, et al. Prevalence and genetic diversity of human caliciviruses (HuCVs) in Mexican children. J Med Virol. 2000;62:217–23.PubMedCrossRef
14.
go back to reference Fasel D, Mellmann A, Cernela N, et al. Hemolytic uremic syndrome in a 65 year-old male linked to a very unusual type of stx2e and eae harboring O51:H49 Shiga-toxin producing Escherichia coli. J Clin Microbiol. 2014;52:1301–3.PubMedCentralPubMedCrossRef Fasel D, Mellmann A, Cernela N, et al. Hemolytic uremic syndrome in a 65 year-old male linked to a very unusual type of stx2e and eae harboring O51:H49 Shiga-toxin producing Escherichia coli. J Clin Microbiol. 2014;52:1301–3.PubMedCentralPubMedCrossRef
15.
go back to reference Flem ET, Latipov R, Nurmatov ZS, et al. Costs of diarrheal disease and the cost-effectiveness of a rotavirus vaccination program in kyrgyzstan. J Infect Dis. 2009;200:S195–202.PubMedCrossRef Flem ET, Latipov R, Nurmatov ZS, et al. Costs of diarrheal disease and the cost-effectiveness of a rotavirus vaccination program in kyrgyzstan. J Infect Dis. 2009;200:S195–202.PubMedCrossRef
16.
go back to reference Henriksen SA, Pohlenz JF. Staining of cryptosporidia by a modified Ziehl-Neelsen technique. Acta Vet Scand. 1981;22:594–6.PubMed Henriksen SA, Pohlenz JF. Staining of cryptosporidia by a modified Ziehl-Neelsen technique. Acta Vet Scand. 1981;22:594–6.PubMed
17.
go back to reference Jiang ZD, Lowe B, Verenkar MP, et al. Prevalence of enteric pathogens among international travelers with diarrhea acquired in Kenya (Mombasa), India (Goa), or Jamaica (Montego Bay). J Infect Dis. 2002;185:497–502.PubMedCrossRef Jiang ZD, Lowe B, Verenkar MP, et al. Prevalence of enteric pathogens among international travelers with diarrhea acquired in Kenya (Mombasa), India (Goa), or Jamaica (Montego Bay). J Infect Dis. 2002;185:497–502.PubMedCrossRef
18.
go back to reference Johnson S, Gerding DN. Clostridium difficile—associated diarrhea. Clin Infect Dis. 1998;26:1027–34 (quiz 1035–1026).PubMedCrossRef Johnson S, Gerding DN. Clostridium difficile—associated diarrhea. Clin Infect Dis. 1998;26:1027–34 (quiz 1035–1026).PubMedCrossRef
19.
go back to reference Kirkwood CD, Bishop RF. Molecular detection of human calicivirus in young children hospitalized with acute gastroenteritis in Melbourne, Australia, during 1999. J Clin Microbiol. 2001;39:2722–4.PubMedCentralPubMedCrossRef Kirkwood CD, Bishop RF. Molecular detection of human calicivirus in young children hospitalized with acute gastroenteritis in Melbourne, Australia, during 1999. J Clin Microbiol. 2001;39:2722–4.PubMedCentralPubMedCrossRef
20.
go back to reference Laubach HE, Bentley CZ, Ginter EL, et al. A study of risk factors associated with the prevalence of Cryptosporidium in villages around Lake Atitlan, Guatemala. Braz J Infect Dis. 2004;8:319–23.PubMedCrossRef Laubach HE, Bentley CZ, Ginter EL, et al. A study of risk factors associated with the prevalence of Cryptosporidium in villages around Lake Atitlan, Guatemala. Braz J Infect Dis. 2004;8:319–23.PubMedCrossRef
21.
go back to reference Lindell AT, Grillner L, Svensson L, et al. Molecular epidemiology of norovirus infections in Stockholm, Sweden, during the years 2000 to 2003: association of the GGIIb genetic cluster with infection in children. J Clin Microbiol. 2005;43:1086–92.PubMedCentralPubMedCrossRef Lindell AT, Grillner L, Svensson L, et al. Molecular epidemiology of norovirus infections in Stockholm, Sweden, during the years 2000 to 2003: association of the GGIIb genetic cluster with infection in children. J Clin Microbiol. 2005;43:1086–92.PubMedCentralPubMedCrossRef
22.
go back to reference Liu L, Johnson HL, Cousens S, et al. Global, regional, and national causes of child mortality: an updated systematic analysis for 2010 with time trends since 2000. Lancet. 2012;379:2151–61.PubMedCrossRef Liu L, Johnson HL, Cousens S, et al. Global, regional, and national causes of child mortality: an updated systematic analysis for 2010 with time trends since 2000. Lancet. 2012;379:2151–61.PubMedCrossRef
23.
go back to reference Marie-Cardine A, Gourlain K, Mouterde O, et al. Epidemiology of acute viral gastroenteritis in children hospitalized in Rouen, France. Clin Infect Dis. 2002;34:1170–8.PubMedCrossRef Marie-Cardine A, Gourlain K, Mouterde O, et al. Epidemiology of acute viral gastroenteritis in children hospitalized in Rouen, France. Clin Infect Dis. 2002;34:1170–8.PubMedCrossRef
24.
go back to reference Marti H, Escher E. SAF—an alternative fixation solution for parasitological stool specimens. Schweiz Med Wochenschr. 1990;120:1473–6.PubMed Marti H, Escher E. SAF—an alternative fixation solution for parasitological stool specimens. Schweiz Med Wochenschr. 1990;120:1473–6.PubMed
25.
go back to reference Martin-Ampudia M, Mariscal A, Lopez-Gigosos RM, et al. Under-notification of cryptosporidiosis by routine clinical and laboratory practices among non-hospitalised children with acute diarrhoea in Southern Spain. Infection. 2012;40:113–9.PubMedCrossRef Martin-Ampudia M, Mariscal A, Lopez-Gigosos RM, et al. Under-notification of cryptosporidiosis by routine clinical and laboratory practices among non-hospitalised children with acute diarrhoea in Southern Spain. Infection. 2012;40:113–9.PubMedCrossRef
26.
go back to reference Meinhardt PL, Casemore DP, Miller KB. Epidemiologic aspects of human cryptosporidiosis and the role of waterborne transmission. Epidemiol Rev. 1996;18:118–36.PubMedCrossRef Meinhardt PL, Casemore DP, Miller KB. Epidemiologic aspects of human cryptosporidiosis and the role of waterborne transmission. Epidemiol Rev. 1996;18:118–36.PubMedCrossRef
27.
go back to reference Mengelle C, Mansuy JM, Prere MF, et al. Simultaneous detection of gastrointestinal pathogens with a multiplex Luminex-based molecular assay in stool samples from diarrhoeic patients. Clin Microbiol Infect. 2013;19:E458–65.PubMedCrossRef Mengelle C, Mansuy JM, Prere MF, et al. Simultaneous detection of gastrointestinal pathogens with a multiplex Luminex-based molecular assay in stool samples from diarrhoeic patients. Clin Microbiol Infect. 2013;19:E458–65.PubMedCrossRef
28.
go back to reference Moore JE, Madden RH. Survival of Campylobacter coli in porcine liver. Food microbiology. New York: Academic Press; 2001. p. 1–10. Moore JE, Madden RH. Survival of Campylobacter coli in porcine liver. Food microbiology. New York: Academic Press; 2001. p. 1–10.
29.
go back to reference Morin N, Tirling C, Ivison SM, et al. Autoactivation of the AggR regulator of enteroaggregative Escherichia coli in vitro and in vivo. FEMS Immunol Med Microbiol. 2010;58:344–55.PubMed Morin N, Tirling C, Ivison SM, et al. Autoactivation of the AggR regulator of enteroaggregative Escherichia coli in vitro and in vivo. FEMS Immunol Med Microbiol. 2010;58:344–55.PubMed
30.
go back to reference Murray PR, Boron EJ, Jorgensen JH, Landry ML, Pfaller MA. Manual of clinical microbiology. Washington: American Society of Microbiology Press; 2007. Murray PR, Boron EJ, Jorgensen JH, Landry ML, Pfaller MA. Manual of clinical microbiology. Washington: American Society of Microbiology Press; 2007.
32.
go back to reference Owen RJ, On SL, Costas M. The effect of cooling rate, freeze-drying suspending fluid and culture age on the preservation of Campylobacter pylori. J Appl Bacteriol. 1989;66:331–7.PubMedCrossRef Owen RJ, On SL, Costas M. The effect of cooling rate, freeze-drying suspending fluid and culture age on the preservation of Campylobacter pylori. J Appl Bacteriol. 1989;66:331–7.PubMedCrossRef
33.
go back to reference Rheingans R, Kukla M, Adegbola RA, et al. Exploring household economic impacts of childhood diarrheal illnesses in three African settings. Clin Infect Dis. 2012;55:S317–26.PubMedCentralPubMedCrossRef Rheingans R, Kukla M, Adegbola RA, et al. Exploring household economic impacts of childhood diarrheal illnesses in three African settings. Clin Infect Dis. 2012;55:S317–26.PubMedCentralPubMedCrossRef
34.
go back to reference Roman E, Wilhelmi I, Colomina J, et al. Acute viral gastroenteritis: proportion and clinical relevance of multiple infections in Spanish children. J Med Microbiol. 2003;52:435–40.PubMedCrossRef Roman E, Wilhelmi I, Colomina J, et al. Acute viral gastroenteritis: proportion and clinical relevance of multiple infections in Spanish children. J Med Microbiol. 2003;52:435–40.PubMedCrossRef
35.
go back to reference Scheier E, Aviner S. Septicemia following rotavirus gastroenteritis. Isr Med Assoc J. 2013;15:166–9.PubMed Scheier E, Aviner S. Septicemia following rotavirus gastroenteritis. Isr Med Assoc J. 2013;15:166–9.PubMed
36.
go back to reference Schmidt H, Knop C, Franke S, et al. Development of PCR for screening of enteroaggregative Escherichia coli. J Clin Microbiol. 1995;33:701–5.PubMedCentralPubMed Schmidt H, Knop C, Franke S, et al. Development of PCR for screening of enteroaggregative Escherichia coli. J Clin Microbiol. 1995;33:701–5.PubMedCentralPubMed
37.
go back to reference Shkalim V, Amir A, Samra Z, et al. Characteristics of non-typhi Salmonella gastroenteritis associated with bacteremia in infants and young children. Infection. 2012;40:285–9.PubMedCrossRef Shkalim V, Amir A, Samra Z, et al. Characteristics of non-typhi Salmonella gastroenteritis associated with bacteremia in infants and young children. Infection. 2012;40:285–9.PubMedCrossRef
38.
go back to reference Simpson R, Aliyu S, Iturriza-Gomara M, et al. Infantile viral gastroenteritis: on the way to closing the diagnostic gap. J Med Virol. 2003;70:258–62.PubMedCrossRef Simpson R, Aliyu S, Iturriza-Gomara M, et al. Infantile viral gastroenteritis: on the way to closing the diagnostic gap. J Med Virol. 2003;70:258–62.PubMedCrossRef
39.
go back to reference Stark D, Al-Qassab SE, Barratt JL, et al. Evaluation of multiplex tandem real-time PCR for detection of Cryptosporidium spp., Dientamoeba fragilis, Entamoeba histolytica, and Giardia intestinalis in clinical stool samples. J Clin Microbiol. 2011;49:257–62.PubMedCentralPubMedCrossRef Stark D, Al-Qassab SE, Barratt JL, et al. Evaluation of multiplex tandem real-time PCR for detection of Cryptosporidium spp., Dientamoeba fragilis, Entamoeba histolytica, and Giardia intestinalis in clinical stool samples. J Clin Microbiol. 2011;49:257–62.PubMedCentralPubMedCrossRef
40.
go back to reference Thielman NM, Guerrant RL. Clinical practice. Acute infectious diarrhea. N Engl J Med. 2004;350:38–47.PubMedCrossRef Thielman NM, Guerrant RL. Clinical practice. Acute infectious diarrhea. N Engl J Med. 2004;350:38–47.PubMedCrossRef
Metadata
Title
Gastrointestinal pathogens detected by multiplex nucleic acid amplification testing in stools of pediatric patients and patients returning from the tropics
Authors
C. Beckmann
U. Heininger
H. Marti
H. H. Hirsch
Publication date
01-12-2014
Publisher
Springer Berlin Heidelberg
Published in
Infection / Issue 6/2014
Print ISSN: 0300-8126
Electronic ISSN: 1439-0973
DOI
https://doi.org/10.1007/s15010-014-0656-7

Other articles of this Issue 6/2014

Infection 6/2014 Go to the issue
Live Webinar | 27-06-2024 | 18:00 (CEST)

Keynote webinar | Spotlight on medication adherence

Live: Thursday 27th June 2024, 18:00-19:30 (CEST)

WHO estimates that half of all patients worldwide are non-adherent to their prescribed medication. The consequences of poor adherence can be catastrophic, on both the individual and population level.

Join our expert panel to discover why you need to understand the drivers of non-adherence in your patients, and how you can optimize medication adherence in your clinics to drastically improve patient outcomes.

Prof. Kevin Dolgin
Prof. Florian Limbourg
Prof. Anoop Chauhan
Developed by: Springer Medicine
Obesity Clinical Trial Summary

At a glance: The STEP trials

A round-up of the STEP phase 3 clinical trials evaluating semaglutide for weight loss in people with overweight or obesity.

Developed by: Springer Medicine

Highlights from the ACC 2024 Congress

Year in Review: Pediatric cardiology

Watch Dr. Anne Marie Valente present the last year's highlights in pediatric and congenital heart disease in the official ACC.24 Year in Review session.

Year in Review: Pulmonary vascular disease

The last year's highlights in pulmonary vascular disease are presented by Dr. Jane Leopold in this official video from ACC.24.

Year in Review: Valvular heart disease

Watch Prof. William Zoghbi present the last year's highlights in valvular heart disease from the official ACC.24 Year in Review session.

Year in Review: Heart failure and cardiomyopathies

Watch this official video from ACC.24. Dr. Biykem Bozkurt discusses last year's major advances in heart failure and cardiomyopathies.