Molecular Prevalence of Entamoeba Species among Diarrheal Patients in Eastern Kenya



Abstract Views
455

Downloads
312

Citations

Share

##plugins.themes.bootstrap3.article.sidebar##

Submitted Nov 5, 2021
Published Jan 24, 2022
10.24018/ejbio.2022.3.1.304

Abstract viewed = 455 times

Table of Contents

##plugins.themes.bootstrap3.article.main##

Data on the epidemiology of Entamoeba infections in Eastern part of Kenya is scanty. Diagnostic tests in use have limited capacity to differentiate common infecting species. The performance of Polymerase Chain Reaction (PCR) to differentiate between pathogenic Entamoeba histolytica and two nonpathogenic species E. dispar and E. moshkovskii is largely undetermined. Therefore, this study sought to determine the prevalence and associated factors for Entamoeba infections and evaluate the performance of PCR to differentiate Entamoeba complex species among diarrheal patient in Eastern region of Kenya. Stool samples were obtained from 400 patients attending Meru Teaching and Referral Hospital (MTRH) presenting with diarrhea. Samples were processed by direct wet mount using normal saline and iodine stain for microscopic examination. Entamoeba species differentiation was done using PCR targeting the 16S rRNA gene. A total of 33 (8.3%) samples had Entamoeba cysts/trophozoites by microscopy while 29 (7.3%) were identified as E. histolytica by PCR. Entamoeba infections was most common among adults 23 (5.8%) and in females 20(5%). The sensitivity of microscopy was 29/29 (100%; 95% CI 88.1% - 100%) with a specificity 367/371 (98.9%; 95% CI 97.3 % - 99.7%). In multivariate analysis, factors that independently influenced Entamoeba infection included sources of drinking water, use of toilet with water, regular use of soap or sanitizer, having diarrhea that persists for two weeks and stool consistency. Entamoeba infection was found to be responsible for most diarrhea condition especially among children. Patients hygienic and sanitation characteristics contributes significantly to Entamoeba infection. The performance of microscopy to detect Entamoeba infection is comparable to those of PCR except for the lack of species differentiation. Molecular species differentiation will improve disease diagnosis, control and management. Continuous monitoring of patient presenting with diarrhea for Entamoeba infection would improve treatment outcomes.

Keywords: Diarrheal diseases, Eastern Kenyan, Entamoeba species, Entamoeba dispar, Entamoeba moshkovskii, epidemiology, molecular diagnostics

References

  1. Abu-Madi, M., Boughattas, S., Behnke, J. M., Sharma, A., Ismail, A. (2017). Coproscopy and molecular screening for detection of intestinal protozoa. Parasit Vectors, 10.
     Google Scholar
  2. Al-Harthi, S., Jamjoom, M. (2007). Diagnosis and Differentiation of Entamoeba infection in Makhah Al Mukarramah using microscopy and stool Antigen Detection Kits. W J Med Sci, 2: 15-20.
     Google Scholar
  3. Al‐Areeqi, M. A., Sady, H., Al‐Mekhlafi, H., Anuar, T., Al‐Adhroey, A., Atroosh, W., et al (2017). First molecular epidemiology of Entamoeba histolytica, E. dispar and E. moshkovskii infections in Yemen: different species‐specific associated risk factors. Tropical Medicine and International Health. 22: 493-504.
     Google Scholar
  4. Aly, N. S., Bayoumi, I., Selem, R., Kardoush, M., Rashed, G., Moharam, A. (2018). A Novel Nano Magnetic Beads Dot ELISA Immunoassay and Its Application on the Detection of Giardia lamblia Coproanti-gen. Iran J Parasitol, 13(4): 532–40.
     Google Scholar
  5. Anuar, T. S., Al-Mekhlafi, H. M., Ghani, M. K. A., Bakar, E. A., Azreen, S. N., Salleh, F. M., et al. (2012). Molecular epidemiology of amoebiasis in Malaysia: highlighting the different risk factors of Entamoeba histolytica and Entamoeba dispar infections among Orang Asli communities. International Journal of Parasitology, 42: 1165 – 1175.
     Google Scholar
  6. Begum, S., Quach, J., & Chadee, K. (2015). Immune evasion mechanisms of Entamoeba histolytica: progression to disease. Frontiers in microbiology, 6, 1394.
     Google Scholar
  7. Benetton, M. L., Goncalves, A. V., Meneghini, M. E., Silva, E. F., Carneiro, M. (2005). Risk factors for infection by the Entamoeba histolytica/E. dispar complex: an epidemiological study conducted in outpatient clinics in the city of Manaus, Amazon Region, Brazil. Trans R Soc Trop Med Hyg, 99: 532-540.
     Google Scholar
  8. Center for Disease Control and Prevention. (2016). Global Water, Sanitation & Hygiene (WASH).
     Google Scholar
  9. Centre for Disease Control and Prevention (2015). Parasites - Amebiasis -Entamoeba Histolytica Infection.
     Google Scholar
  10. Cheesbrough, M. (2012). District Laboratory Practice in Tropical Countries. Cambridge University Press.
     Google Scholar
  11. Development Initiative organization (2018). Enhancing access to safe water and improved sanitation services in Kenya (Issue December).
     Google Scholar
  12. Duc, P. P., Nguyen-Viet, H., Hattendorf, J., Zinnstag, J., Cam, P., Odermatt, P. (2011). Risk factors for Entamoeba histolytica infection in an agricultural community in Hanam province, Vietnam. Parasites Vectors, 4: 102.
     Google Scholar
  13. Easton, A. V., Oliveira, R. G., O’Connell, E. M., Kepha, S., Mwandawiro, C. S., Njenga, S. M., et al. (2016). Multi-parallel qPCR provides increased sensitivity and diagnostic breadth for gastrointestinal parasites of humans: field-based inferences on the impact of mass deworming. Parasites Vectors, 9: 38
     Google Scholar
  14. Elston, D. M. (2006). Laboratory Results That Should Be Ignored. Med Gen Med, 8(4): 9.
     Google Scholar
  15. Emisiko, J., Shavia, N., Shiluli, C., Kiboi, N., Wamalwa, R., Jumba, B., et al. (2020). Comparison of Microscopy and PCR for Detection of Giardia Lamblia and Entamoeba Histolytica in Human Stool Specimens in a Resource Limited Setting in Western Kenya. Ethiop J Health Sci. 30(6).
     Google Scholar
  16. 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. J Clin Microbiol, 45(3): 1035–7.
     Google Scholar
  17. Hamzah, Z., Petmitr, S., Mungthin, M., Leelayoova, S., Chavalitshewinkoon- Petmitr, P. (2010). Development of multiplex real-time polymerase chain reaction for detection of Entamoeba histolytica, Entamoeba dispar and Entamoeba moshkovskii in clinical specimens. Am J Trop Med Hyg, 83: 909–913.
     Google Scholar
  18. Haque, R., D. Mondal, B. D. Kirkpatrick, Akther, S., Farr, B. M., Sack, R. B., Petri, Jr., W. A. (2003). Epidemiologic and clinical characteristics of acute diarrhea with emphasis on E. histolytica infections in preschool children in urban slum of Dhaka, Bangladesh. Am. J. Trop. Med. Hyg, 69: 398-405.
     Google Scholar
  19. Kagira, J. M., Maina, N., Njenga, J., Karanja, S. M., Karori, S. M., Ngotho, J. M. (2011). Prevalence and types of coinfections in sleeping sickness patients in Kenya (2000/2009). J Trop Med.
     Google Scholar
  20. 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 Microbiol, 7: 47.
     Google Scholar
  21. Kosek, M., Bern, C., Guerrant, R. L. (2003). The global burden of diarrhoeal disease, as estimated from studies published between 1992 and 2000. Bull, 81: 197-20.
     Google Scholar
  22. Lau, Y. L., Anthony, C., Fakhrurrazi, S. A., Ibrahim, J., Ithoi, I., Mahmud, R. (2013). Real-time PCR assay in differentiating Entamoeba histolytica, Entamoeba dispar, and Entamoeba moshkovskii infections in Orang Asli settlements in Malaysia. Parasites Vectors, 6: 250.
     Google Scholar
  23. Lemeshow, S., Hosmer, D.K., Klar, J., Lwanga, S.K., World Heath Organization. (1990). World health Organization. Adequacy of samples size in health studies. Chichester: Wiley.
     Google Scholar
  24. López-López, P., Martínez-López, M. C., Boldo-León, X. M., Hernández-Díaz, Y., Gonzalez- Castro, T. B., Tovilla-Zarate, C., et al., (2017). Detection and differentiation of Entamoeba histolytica and Entamoeba dispar in clinical samples through PCR-denaturing gradient gel electrophoresis. Brazilian Journal of Medical and Biological Research, 50(4), 1–7.
     Google Scholar
  25. Matey, E. J., Tokoro, M., Nagamoto, T., Mizuno, T., Saina, M. C., Xiuqiong, B., et al. (2016). Lower prevalence of Entamoeba species in children with vertically transmitted HIV infection in Western Kenya. AIDS, 30: 803–5.
     Google Scholar
  26. Mbae, C. K., Nokes, D. J., Mulinge, E., Nyambura, J., Waruru, A., Kariuki, S. (2013). Intestinal parasitic infections in children presenting with diarrhoea in outpatient and inpatient settings in an informal settlement of Nairobi, Kenya. BMC Infect Dis, 13: 243.
     Google Scholar
  27. Mulinge, E., Mbae, C., Ngugi, B., Irungu, T., Matey, E., & Kariuki, S. (2021). Entamoeba species infection in patients seeking treatment for diarrhea and abdominal discomfort in Mukuru informal settlement in Nairobi, Kenya. Food and Waterborne Parasitology, 23, e00122.
     Google Scholar
  28. Norhayati, M., Fatmah, M. S., Yusof, S., Edariah, A. B. (2003). Intestinal parasitic infections in man: a review. Med J Malaysia, 58: 296-305.
     Google Scholar
  29. Nowak, P. (2015). Entamoeba histolytica - Pathogenic Protozoan of the Large Intestine in Humans. Journal of Clinical Microbiology and Biochemical Technology, 1, 010–017.
     Google Scholar
  30. Parija, S. C., Mandal, J., Ponnambath, D. K. (2014). Laboratory methods of identification of Entamoeba histolytica and its differentiation from look-alike Entamoeba spp. Tropical Parasitology, 4(2): 90.
     Google Scholar
  31. Pereira, V. V., Conceição, A. S., Maximiano, L. H. S., Belligoli, L. Q. G., Silva, E. S., Pereira, V. V. (2014). Laboratory diagnosis of amebiasis in a sample of students from southeastern Brazil and a comparison of microscopy with enzyme-linked immunosorbent assay for screening of infections with Entamoeba sp. Revista da Sociedade Brasileira de Medicina Tropical. 47(1):52
     Google Scholar
  32. Petri, W. A., Haque, R., Lyerly, D., Vines, R. (2000). Estimating the impact of amoebiasis on health. Parasitol Today, 16: 320-321.
     Google Scholar
  33. Qvarnstrom, Y., James, C., Xayavong, M., Holloway, B. P., Visvesvara, G. S., Sriram, R., et al. (2005). Comparison of real-time PCR protocols for differential laboratory diagnosis of Amebiasis. J Clin Microbiol, 43: 5491–5497.
     Google Scholar
  34. Roberts, L., Janovy Jr. J. (2009). Foundation of Parasitology McGraw-Hill: New York.
     Google Scholar
  35. Sebastian, H. J. V., Stark, D. J., Fotedar, R., Marriott, D., John, T. E. J. T., Harkness, J.L (2007). Amoebiasis: current status in Australia. M J A, 186: 412-416.
     Google Scholar
  36. Shirley, D.-A. T., Farr, L., Watanabe, K., Moonah, S. (2018). A Review of the Global Burden, New Diagnostics, and Current Therapeutics for Amebiasis. Open Forum Infect Dis, 5(7): ofy161.
     Google Scholar
  37. Stauffer, W., Abdalla, M., Ravdin, J. (2006). Prevalence and Incidence of Entamoeba histolytica Infection in South Africa and Egypt. Archives of Medical Research. 37: 265-268.
     Google Scholar
  38. Tanyuksel, M., Petri, W. A. (2003). Laboratory diagnosis of Amebiasis. Clin Microbiol Rev, 16: 713-729.
     Google Scholar
  39. World Health Organization. (2014). Progress on Drinking Water and Sanitation.
     Google Scholar
  40. World Health Organization. (2019). Water sanitation hygiene.
     Google Scholar
  41. Zebardast, N., Haghighi, A., Yeganeh, F., Tabaei, S. J. S., Gharavi, M. J., Fallahi S., et al., (2014). Application of Multiplex PCR for Detection and Differentiation of Entamoeba histolytica, Entamoeba dispar and Entamoeba moshkovskii. Iranian J Parasitol, 466–473.
     Google Scholar