ACUTE ENTERIC INFECTIONS POLYMERASE CHAIN REACTION ASSAY IN PEDIATRIC PRACTICE: OPPORTUNITIES AND CHALLENGES

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Abstract

The aim of the study is estimate the opportunities of local multi-prime PCR reagents kits in children enteric infections etiological diagnostics amongst the patients with diarrhoea vs traditional bacteriological methods. We used 4 kits of reagents that provide multiple pathogens simultaneous indication in one sample: 1) Rotavirus, Norovirus, Astrovirus; 2) Shigella spp./EIEC, Salmonella spp., Campylobacter spp.; 3) Yersinia enterocolitica and Yersinia pseudotuberculosis; 4) E. coli: EIEC (enteroinvasive), EPEC (enteropathogenic), ETEC (enterotoxigenic), EHEC (enterohaemorrhagic), EAgEC (enteroaggregative). It has been shown that the viral intestinal infections is increased by 14%, bacterial — in 2,5 times. PCR diagnostics identified in 62% of patients the viral gastroenteritis: Rotavirus (52%), Norovirus (9%), Astrovirus (1%). Detected bacterial pathogens PCR markers number proved up to 2.5 times high than according to bacteriological examination. The spectrum of bacterial agents increased due to E. coli and Y. enterocolitica. PCR diagnostics increased detection of Campylobacter up to 2 times. Detected E. coli DNA prevalence: EPEC — 66%, EAgEC, ETEC and EHEC were 31%, 9% and 4%, respectively. DNA Campylobacter spp. and E. coli constituted 2/3 of all findings: Campylobacter spp. (41%), E. coli (24%), Salmonella spp. (19%), Yersinia spp. (11%), Shigella spp./EIEC (5%). The positive results of bacteriological and serological methods duplicate the positive results of PCR diagnostics. In general, the positive results of PCR diagnosis of bacterial pathogens were detected in 46.35% of the examined patients. In 48.4% of patients identified PCR markers viral — bacterial infection, in 5.25% — of bacterial associations, in 11% of them were found the DNA 2–3 bacterial pathogens. The study was shown in children in St. Petersburg in 2012–2014 dominated rotavirus infection, campylobacteriosis and escherichiosis. The prevalence of viral-bacterial confections is more than 50% of all diagnosed cases.

About the authors

E. D. Sokolova

St. Petersburg Children’s Municipal Clinical Hospital named after N.F. Filatov, St. Petersburg, Russian Federation

Author for correspondence.
Email: ed_sokolova@mail.ru
PhD, MD (Biology), Senior Researcher, Virologist, Clinical Diagnostic Laboratory, Children’s Municipal Clinical Hospital named after N.F. Filatov, St. Petersburg, Russian Federation Russian Federation

A. M. Galtaeva

St. Petersburg Children’s Municipal Clinical Hospital named after N.F. Filatov, St. Petersburg, Russian Federation

Email: fake@neicon.ru
Bacteriologist, Clinical Diagnostic Laboratory, Children’s Municipal Clinical Hospital named after N.F. Filatov, St. Petersburg, Russian Federation Russian Federation

O. U. Zamurei

St. Petersburg Children’s Municipal Clinical Hospital named after N.F. Filatov, St. Petersburg, Russian Federation

Email: fake@neicon.ru
Bacteriologist, Clinical Diagnostic Laboratory, Children’s Municipal Clinical Hospital named after N.F. Filatov, St. Petersburg, Russian Federation Russian Federation

O. V. Didichenko

St. Petersburg Children’s Municipal Clinical Hospital named after N.F. Filatov, St. Petersburg, Russian Federation

Email: fake@neicon.ru
Bacteriologist, Clinical Diagnostic Laboratory, Children’s Municipal Clinical Hospital named after N.F. Filatov, St. Petersburg, Russian Federation Russian Federation

U. V. Sokolova

St. Petersburg Children’s Municipal Clinical Hospital named after N.F. Filatov, St. Petersburg, Russian Federation

Email: fake@neicon.ru
Laboratory Diagnostics Specialist, Clinical Diagnostic Laboratory, Children’s Municipal Clinical Hospital named after N.F. Filatov, St. Petersburg, Russian Federation Russian Federation

V. A. Muratova

St. Petersburg Children’s Municipal Clinical Hospital named after N.F. Filatov, St. Petersburg, Russian Federation

Email: fake@neicon.ru
Bacteriologist, Clinical Diagnostic Laboratory, Children’s Municipal Clinical Hospital named after N.F. Filatov, St. Petersburg, Russian Federation Russian Federation

O. U. Ligorova

St. Petersburg Children’s Municipal Clinical Hospital named after N.F. Filatov, St. Petersburg, Russian Federation

Email: fake@neicon.ru
Bacteriologist, Clinical Diagnostic Laboratory, Children’s Municipal Clinical Hospital named after N.F. Filatov, St. Petersburg, Russian Federation Russian Federation

I. N. Zhuravleva

St. Petersburg Children’s Municipal Clinical Hospital named after N.F. Filatov, St. Petersburg, Russian Federation

Email: fake@neicon.ru
Bacteriologist, Clinical Diagnostic Laboratory, Children’s Municipal Clinical Hospital named after N.F. Filatov, St. Petersburg, Russian Federation Russian Federation

M. A. Makarova

St. Petersburg Pasteur Institute, St. Petersburg, Russian Federation

Email: fake@neicon.ru
PhD (Medicine), Senior Researcher, Laboratory of Enteric Infections, St. Petersburg Pasteur Institute, St. Petersburg, Russian Federation Russian Federation

L. A. Kaftyreva

St. Petersburg Pasteur Institute, St. Petersburg, Russian Federation

Email: fake@neicon.ru
PhD, MD (Medicine), Professor, Head of the Laboratory of Enteric Infections, St. Petersburg Pasteur Institute, St. Petersburg, Russian Federation Russian Federation

References

  1. Бабик Р.Л., Сагалова О.И. Оптимизация диагностики вирусных и бактериальных кишечных инфекций у детей и взрослых // Инфекционные болезни. 2015. Т. 13, № 2. С. 46–54. [Babik R.K., Sagalova O.I. Optimization of diagnostics of viral and bacterial enteric infections in children and adolescents. Infektsionnye bolezni = Infectious Diseases, 2015, vol. 13, no. 2, pp. 46–54 (In Russ.)]
  2. Горелов А.В., Бондарева А.В., Подколзин А.Т. Клинико-эпидемиологическая характеристика энтероаггрегативного эшерихиоза у детей // Инфекционные болезни. 2013. Т. 11, № 3. С. 22–26. [Gorelov A.V., Bondareva A.V., Podkolzin A.T. Clinical and epidemiology feathes of enteroaggregative escherichia infection in children. Infektsionnye bolezni = Infectious Diseases, 2013, vol. 11, no. 3, pp. 22–26 (In Russ.)]
  3. О состоянии санитарно-эпидемиологического благополучия населения в Российской Федерации в 2014 году: Государственный доклад Федеральной службы по надзору в сфере защиты прав потребителей и благополучия человека, 2015. 206 c. [On the state sanitary and epidemiological welfare of the population in the Russian Federation in 2014: State report]. 2015. 206 p.]. URL: http://rospotrebnadzor.ru/upload/iblock/22c/gd_2014_seb_dlya-sayta.pdf (дата обращения: 14.06.2016).
  4. Сведения об инфекционных и паразитарных заболеваниях (форма 1) за январь-декабрь 2012, 2013, 2014 гг. [Information about infectious and parasitic diseases (Form 1) for January-December 2012, 2013, 2014.]. URL: http://rospotrebnadzor.ru/activities/statistical-materials/?type=special (дата обращения: 14.06.2016).
  5. Antikainen J., Kantele A., Pakkanen S.H., Laaveri T., Riutta J., Vaara M., Kirveskari J. A quantitative polymerase chain reaction assay for rapid detection of 9 pathogens directly from stools of travelers with diarrhea. Clin. Gastroenterol. Hepatol., 2013, vol. 11, iss. 10, pp. 1300–1307. doi: 10.1016/j.cgh.2013.03.037
  6. Barletta F., Ochoa T.J., Mercado E., Ruiz J., Ecker L., Lopez G., Mispireta M., Gil A.I., Lanata C.F., Cleary T.G. Quantitative real-time polymerase chain reaction for enteropathogenic Escherichia coli: a tool for investigation of asymptomatic versus symptomatic infections. Clin. Infect. Dis., 2011, vol. 53, no. 12, pp. 1223–1229. doi: 10.1093/cid/cir730
  7. Becker S.L., Chatiqre J.K., Gohou J.P., Coulibaly J.T., Leuppi R., Polman K., Chappuis F., Mertens P., Herrman M., N’Goran E.K., Utzinqer J., Von Muller L. Combained stool-based multiplex PCR and microscopy for enhanced pathogen detection in patients with persistent diarrhea and asymptomatic controls from Cote d’Ivoire. Clin. Microbiol. Infect., 2015, vol. 21, no. 6: 591. doi: 10.1016/j.cmi.2015.02.016
  8. David E.B., Guimaraes S., De Oliveira A.P., Goulart de Oliveira-Sequeria T.C., Noqueira Bittencourt G., Moraes Nardi A.R., Martins Pibolla P.E., Bueno-Franco R.M., Branco N., Tosini F., Bella A., Pozio E., Caccio S.M. Molecular characterization of intestinal protozoa in two poor communities in the State of Sao Paulo, Brasil. Parasit. Vectors, 2015, vol. 8: 103. doi: 10.1186/s13071-015-0714-8
  9. Goldfarb D.M., Dixon B., Moldovan I., Barrowman N., Mattison K., Zentner C., Baikie M., Bidowid S., Chan F., Slinqer R. Nanolitre real-time PCR detection of bacterial, parasitic, and viral agents from patients with diarrhea in Nunavut, Canada. Int. J. Circumpolar Health, 2013, vol. 72: 19903. doi: 10.3402/ijch.v72i0.19903
  10. Laaveri T., Pakkanen S.H., Antikainen J., RiuttaJ., Mero S., Kirveskari J., Antele A. High number of diarrhoeal co-infections in travelers to Benin, West Africa. BMC Infect. Dis., 2014, vol. 14: 81. doi: 10.1186/1471-2334-14-81
  11. Maas L., Dorigo-Zetsma J.W., De Groot C.J., Bouter S., Plotz F.B., Van Ewijk B.E. Detection of intestinal protozoa in paediatric patients with gastrointestinal symptoms by multiplex real-time PCR. Clin. Microbiol. Infect., 2014, vol. 20, no. 6, pp. 545–550. doi: 10.1111/1469-0691.12386
  12. McAuliffe G.N., Anderson T.P., Stevens M., Adams J., Coleman R., Vahagamasera P., Young S., Henderson T., Hoffmann M., Jennings L.C., Murdoch D.R. Systematic application of multiplex PCR enhances the detection of bacteria, parasites and viruses in stool samples. J. Infect., 2013, vol. 67, no. 2, pp. 122–129. doi: 10.1016/j.jinf.2013.04.009
  13. Pankhurst L., Macfarlane-Smith L., Buchanan J., Anson L., Davies K., O’Connor L., Ashwin H., Pike G., Dingle K.E., Peto T.E., Wordsworth S., Walker A.S., Wilcox M.H., Crook D.W. Can rapid integrated polymerase chain reaction-based diagnostics for gastrointestinal pathogens improve routine hospital infection control practice? A diagnostic study. Health Technol. Assess, 2014, vol. 18, no. 53, pp. 1–167. doi: 10.3310/hta18530
  14. Perry M.D., Corden S.A., Howe R.A. Evaluation of the Luminex xTAG Gastrointestinal Pathogen Panel and the Savyon Diagnostics Gastrointestinal Infection Panel for the detection of enteric pathogens in clinical samples. J. Med. Microbiol., 2014, vol. 63, pt. 11, pp. 1419–1426. doi: 10.1099/jmm.0.074773-0
  15. Tennat S.M., Grant T.H., Robins-Browne R.M. Pathogenicity of Yersinia enterocolitica biotype 1A. FEMS Immunol. Med. Microbiol., 2003, vol. 38, no. 2, pp. 127–137.

Copyright (c) 2016 Sokolova E.D., Galtaeva A.M., Zamurei O.U., Didichenko O.V., Sokolova U.V., Muratova V.A., Ligorova O.U., Zhuravleva I.N., Makarova M.A., Kaftyreva L.A.

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