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Licensed Unlicensed Requires Authentication Published by De Gruyter December 28, 2016

Molecular diagnosis of Entamoeba spp. versus microscopy in the Great Cairo

  • Mohamed H. Roshdy , Nour M. Abd El-Kader , Marwa Ali-Tammam , Isabel Fuentes , Magdy M. Mohamed , Nabila A. El-Sheikh and Jose Miguel Rubio EMAIL logo
From the journal Acta Parasitologica

Abstract

Amoebiasis is a human disease produced by Entamoeba histolytica which causes widespread mortality and morbidity worldwide through diarrheal disease and abscess establishment in parenchymal tissues such as liver, lung, and brain. The true prevalence of infection is unknown for most areas of the world due to the difficulty to characterise Entamoeba histolytica versus other non-pathogenic amoebas with identical morphology, as Entamoeba dispar, and Entamoeba moshkovskii. To overcome microscopy misidentification issues, we tested a nested multiplex polymerase chain reaction (PCR) and a real-time PCR on 194 stool samples collected from incoming dysentery patients in Cairo hospitals diagnosed with E. histolytica by microscopy. Nested PCR showed only 20 (10.3%) samples positive to E. histolytica and 17 (8.7%) to E. dispar. The real-time PCR detected only 19 and 11 samples positive to E. histolytica and E. dispar respectively, showing less sensitivity than the nested PCR. The data show that prevalence of E. histolytica in Cairo is lower when specific diagnosis methods are used instead of traditional microscopy, allowing to differentiate between morphologically identical human amoebas species.

Acknowledgements

This work was supported by the Spanish Agency of International Cooperation and Development (AECID) grant number A1/0355539/11 and by the AESI-ISCIII grant number PI14CIII/00014.

References

Ali I.K. 2015. Intestinal amebae. Clinics in Laboratory Medicine,35, 393–422. 10.1016/j.cll.2015.02.009Search in Google Scholar PubMed

Ali I.K., Clark C.G., Petri W.A. 2008. Molecular epidemiology of amebiasis. Infection Genetics and Evolution, 8, 698–70710.1016/j.meegid.2008.05.004Search in Google Scholar PubMed PubMed Central

Banisch D.M., El-Badry A., Klinnert J.V., Ignatius R., El-Dib N. 2015. Simultaneous detection of Entamoeba histolytica/dispar, Giardia duodenalis and cryptosporidia by immunochromatographic assay in stool samples from patients living in the Greater Cairo Region, Egypt. World Journal of Microbiology and Biotechnology, 31, 1251–1258. 10.1007/s11274-015-1875-5Search in Google Scholar PubMed

Bastien P., Procop G.W., Reischl U. 2008. Quantitative Real-Time PCR is not more sensitive than “conventional” PCR. Journal of Clinical Microbiology, 46, 1897–900. 10.1128/JCM.02258-07Search in Google Scholar PubMed PubMed Central

Blessmann J., Buss H., Nu P.A., Dinh B.T., Ngo Q.T., Van A.L., Alla M.D., Jackson T.F., Ravdin J.I., Tannich E. 2002. Real-time PCR for detection and differentiation of Entamoeba histolytica and Entamoeba dispar in fecal samples. Journal of Clinical Microbiology, 40, 4413–441710.1128/JCM.40.12.4413-4417.2002Search in Google Scholar PubMed PubMed Central

Calegar D.A., Nunes B.C., Monteiro K.J., Santos J.P., Toma H.K., Gomes T.F., Lima M.M., Bóia M.N., Carvalho-Costa F.A.2016. Frequency and molecular characterisation of Entamoeba histolytica, Entamoeba dispar, Entamoeba moshkovskii, and Entamoeba hartmanni in the context of water scarcity in northeastern Brazil. Memórias do Instituto Oswaldo Cruz, 111, 114–119. 10.1590/0074-02760150383Search in Google Scholar PubMed PubMed Central

Forsell J., Koskiniemi S., Hedberg I., Edebro H., Evengård B., Granlund M. 2015. Evaluation of factors affecting real-time PCR performance for diagnosis of Entamoeba histolytica and Entamoeba dispar in clinical stool samples. Journal of Medical Microbiology, 64, 1053–1062. 10.1099/jmm.0.000129Search in Google Scholar PubMed

Fotedar R., Stark D., Beebe N., Marriott D., Ellis J., Harkness J. 2007. PCR detection of Entamoeba histolytica, Entamoeba dispar, and Entamoeba moshkovskii in stool samples from Sydney, Australia. Journal of Clinical Microbiology, 45, 1035–103710.1128/JCM.02144-06Search in Google Scholar PubMed PubMed Central

Hamzah Z., Petmitr S., Mungthin M., Leelayoova S., Chavalitshewinkoon-Petmitr P. 2010. Development of multiplex realtime polymerase chain reaction for detection of Entamoeba histolytica, Entamoeba dispar, and Entamoeba moshkovskii in clinical specimens. The American Journal of Tropical Medicine and Hygiene, 83, 909–913. 10.4269/ajtmh.2010.10-0050Search in Google Scholar PubMed PubMed Central

Ibrahim S.S., El-Matarawy O.M., Ghieth M.A., Abu Sarea E.Y., El-Badry A.A. 2015. Copro prevalence and estimated risk of En-tamoeba histolytica in Diarrheic patients at Beni-Suef, Egypt, World Journal of Microbiology and Biotechnology, 31, 385–390. 10.1007/s11274-014-1791-0Search in Google Scholar PubMed

Khairnar K., Parija S.C. 2007. A novel nested multiplex polymerase chain reaction (PCR) assay for differential detection of Entamoeba histolytica, E. moshkovskii and E. dispar DNA in stool samples. BMC Microbiology, 7, 4710.1186/1471-2180-7-47Search in Google Scholar PubMed PubMed Central

Leiva B., Lebbad M., Winiecka-Krusnell J., Altamirano I., Tellez A., Linder E. 2006. Overdiagnosis of Entamoeba histolytica and Entamoeba dispar in Nicaragua: a microscopic, triage parasite panel and PCR study. Archives of Medical Research, 37, 529–53410.1016/j.arcmed.2005.10.009Search in Google Scholar PubMed

Nazeer J.T., El Sayed K.K., von Thien H., El-Sibaei M.M., Abdel Hamid M.Y., Tawfik R.A., Tannich E. 2013. Use of multiplex real-time PCR for detection of common diarrhea causing protozoan parasites in Egypt. Parasitology Research, 112, 595–601. 10.1007/s00436-012-3171-8Search in Google Scholar PubMed

Nazemalhosseini M., Nochi E., Sahebekhtiari N., Rostami Nejad M., Dabiri, H., Zali M.R., Kazemi B., Haghighi A. 2010. Discrimination of Entamoeba moshkovskii in patients with gastrointestinal disorders by single-round PCR. Japanese Journal of Infectious Diseases, 63, 136–13810.7883/yoken.63.136Search in Google Scholar

Sharbatkhori M., Nazemalhosseini M., Cheraghali F., Maghsoodloorad F.S., Taherkhani H., Vakili M. 2014. Discrimination of Entamoeba spp. in children with dysentery. Gastroenterology and Hepatology from Bed to Bench, 7,164–16710.1016/j.clinbiochem.2014.04.072Search in Google Scholar

Skappak C., Akierman S., Belga S., Novak K., Chadee K., Urbanski S.J., Church D., Beck P.L. 2014. Invasive amoebiasis: a review of Entamoeba infections highlighted with case reports. Canadian Journal of Gastroenterology and Hepatology, 28, 355–35910.1155/2014/745130Search in Google Scholar PubMed PubMed Central

Tanyuksel M., Petri W.A. 2003. Laboratory diagnosis of amebiasis. Clinical Microbiology Reviews, 16, 713–72910.1128/CMR.16.4.713-729.2003Search in Google Scholar PubMed PubMed Central

Uslu H., Aktas O., Uyanik M.H. 2016. Comparison of Various Methods in the Diagnosis of Entamoeba histolytica in Stool and Serum Specimens. The Eurasian Journal of Medicine, 48, 124–129. 10.5152/eurasianjmed.2015.0074Search in Google Scholar PubMed PubMed Central

Van den Bossche D., Cnops L., Verschueren J., Van Esbroeck M. 2015. Comparison of four rapid diagnostic tests, ELISA, microscopy and PCR for the detection of Giardia lamblia, Cryptosporidium spp. and Entamoeba histolytica in feces. Journal of Microbiological Methods, 110, 78–84. 10.1016/j.mimet. 2015.01.016Search in Google Scholar

Verweij J.J., Blangé R.A., Templeton K., Schinkel J., Brienen E.A., van Rooyen M.A., van Lieshout L., Polderman A.M. 2004. Simultaneous detection of Entamoeba histolytica, Giardia lamblia, and Cryptosporidium parvum in fecal samples by using multiplex real-time PCR. Journal of Clinical Microbiology, 42, 1220–122310.1128/JCM.42.3.1220-1223.2004Search in Google Scholar PubMed PubMed Central

Zebardast N., Yeganeh F., Gharavi M.J., Abadi A., Seyyed Tabaei S.J., Haghighi A. 2016. Simultaneous detection and differentiation of Entamoeba histolytica, E. dispar, E. moshkovskii, Giardia lamblia and Cryptosporidium spp. in human fecal samples using multiplex PCR and qPCR-MCA. Acta Tropica,162, 233–238. 10.1016/j.actatropica.2016.07.004Search in Google Scholar PubMed

Received: 2016-9-14
Revised: 2016-11-4
Accepted: 2016-11-10
Published Online: 2016-12-28
Published in Print: 2017-3-1

© 2017 W. Stefański Institute of Parasitology, PAS

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