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Erschienen 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

verfasst von: C. Beckmann, U. Heininger, H. Marti, H. H. Hirsch

Erschienen in: Infection | Ausgabe 6/2014

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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.
Literatur
1.
Zurück zum Zitat 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.
Zurück zum Zitat 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.
Zurück zum Zitat 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.
Zurück zum Zitat 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.
Zurück zum Zitat 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.
Zurück zum Zitat 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.
Zurück zum Zitat 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.
Zurück zum Zitat 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.
Zurück zum Zitat 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.
Zurück zum Zitat Denno DM, Shaikh N, Stapp JR, et al. Diarrhea etiology in a pediatric emergency department: a case control study. Clin Infect Dis. 2012;55:897–904.PubMedCentralPubMedCrossRef Denno DM, Shaikh N, Stapp JR, et al. Diarrhea etiology in a pediatric emergency department: a case control study. Clin Infect Dis. 2012;55:897–904.PubMedCentralPubMedCrossRef
12.
Zurück zum Zitat Dupont HL. Acute infectious diarrhea in immunocompetent adults. N Engl J Med. 2014;370:1532–40.PubMedCrossRef Dupont HL. Acute infectious diarrhea in immunocompetent adults. N Engl J Med. 2014;370:1532–40.PubMedCrossRef
13.
Zurück zum Zitat 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.
Zurück zum Zitat 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.
Zurück zum Zitat 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.
Zurück zum Zitat 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.
Zurück zum Zitat 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.
Zurück zum Zitat 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.
Zurück zum Zitat 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.
Zurück zum Zitat 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.
Zurück zum Zitat 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.
Zurück zum Zitat 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.
Zurück zum Zitat 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.
Zurück zum Zitat 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.
Zurück zum Zitat 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.
Zurück zum Zitat 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.
Zurück zum Zitat 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.
Zurück zum Zitat 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.
Zurück zum Zitat 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.
Zurück zum Zitat 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.
Zurück zum Zitat 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.
Zurück zum Zitat 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.
Zurück zum Zitat 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.
Zurück zum Zitat 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.
Zurück zum Zitat 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.
Zurück zum Zitat 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.
Zurück zum Zitat 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.
Zurück zum Zitat 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.
Zurück zum Zitat 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
Metadaten
Titel
Gastrointestinal pathogens detected by multiplex nucleic acid amplification testing in stools of pediatric patients and patients returning from the tropics
verfasst von
C. Beckmann
U. Heininger
H. Marti
H. H. Hirsch
Publikationsdatum
01.12.2014
Verlag
Springer Berlin Heidelberg
Erschienen in
Infection / Ausgabe 6/2014
Print ISSN: 0300-8126
Elektronische ISSN: 1439-0973
DOI
https://doi.org/10.1007/s15010-014-0656-7

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