Comparison of direct microscopy and molecular method to detect amoebiasis cases from stool specimen and also identify the Entamoeba species involved in infection: A study of Nepal

R. K. Das(1), R. Paudya(2), S. K. Singh(3), E. Sulistyowati(4), M. Saud(5),
(1) Department of Microbiology, Kathmandu College of Science and Technology, Kathmandu, Nepal, Nepal
(2) Department of Microbiology, Kathmandu College of Science and Technology, Kathmandu, Nepal, Nepal
(3) Faculty of Economic and Business, University Airlangga, Surabaya, Indonesia, Indonesia
(4) Department of Animal Science, Faculty of Agriculture, University of Bengkulu, Indonesia, Indonesia
(5) Department of Sociology, Universitas Airlangga, Surabaya, Indonesia Department of Sociology, International Islamic University Islamabad, Pakistan, Pakistan

Abstract


Direct microscopic examination of stained or unstained wet mount preparations or fixed-stained smears of clinical material can often provide the etiological diagnosis of an infectious process. A total of 266 human stools specimens from children of america by a combination of microscopic examination and molecular method. In molecular method, nested polymerase chain reaction (Nested-PCR) targeting genomic Entamoeba species was used. Stool specimens were collected from Southern Plains of Nepal and analyzed at the Kathmandu Center for Genomics and Research Laboratory. The stool specimens were processed by wet mount method using saline as well as iodine staining and examined via microscopy for the presence of Entamoeba cysts or trophozoites. Furthermore, the stool specimens were characterized using Nested-PCR targeting genomic Entamoeba species. Based on microscopic examination, the overall prevalence of Entamoeba infection was 17.6% (47/266). The PCR results showed that 52 (19.5%) specimens are successfully generated species-specific amplicons. Males (21.7% in PCR) were more commonly infected compared to females (16.6% in PCR). Comparison by age groups show 10-15 years age-group (26.6% in PCR) had higher infection than age-group 5-10 years (16.6%) years and 1-5 years (15.2%). The infection with E. histolytica (100%; 52/266) was the predominant cause of amoebiasis, while the infection with E. dispar and E. moshkovskii was not found. PCR is a more effective method for the identification of Entamoeba infection than microscopy.


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References


Ali, I.K.M., M.B. Hossain, S. Roy, P. Ayeh-Kumi, J.W.A. Petri, R. Haque, and C.G. Clark. 2003. Entamoeba moshkovskii infections in children, Bangladesh. Emerge Infect Disemerging. 9:580–584.

Braga, L., M.L. Gomes, M.W. Silva, C. Paiva, A. Sales, B.J. Mann. 2001. Entamoeba histolytica and Entamoeba dispar infections as detected by monoclonal antibody in an urban slum in Fortaleza, Northeastern Brazil Rev Soc Bras Med Trop.

Chakraborty, P. K. 2001. A text book of Microbiology, 1st edition. New Central Book agency (P) Ltd, Calcutta.

Cross, P. 1985. Health Aspects of Night soil and Sludge Use in Agriculture and Aquaculture-Part I: Existing Practices and Beliefs in the Utilization of Human Excreta. International Reference Center for Waste Disposal (No 04/85), Dubendorf, Switzerland.

Drechsel, P. C. A. Scott, L. Raschid-Sally, A.B. Redwoodand. 2010. Wastewater irrigation and health - Assessing and mitigating risk in low-income countries. Sterling, VA, London: Earthscan.

Estevez, E. G., J.A. Levine, J. Warren. 1983. Intestinal parasites in a remote village in Nepal J Clin Microbiol. 17:160–1

Fotedar, R. D. Stark, N. Beebe, D. Marriott, J. Ellis, and J. Harkness. 2007. PCR detection of Entamoeba histolytica, Entamoeba dispar, and Entamoeba moshkovskii in stool samples from Sydney, Australia. J Clin Microbiol 45:1035-1037.

Gyawali, N., R. Amatya. 2009. Nepal PH Intestinal parasitosis in school going children of Dharan Municipality, Nepal Trop Gastroenterol. 30 (3):145-7.

Hamzah, Z., S. Petmitr, M. Mungthin, S. Leelayoova, P. Chavalitshewinkoon-Petmitr. 2006. Differential detection of Entamoeba histolytica, Entamoeba dispar, and Entamoeba moshkovskii by a single-round PCR assay J Clin Microbiol. 44:3196–3200.

Haque, R., C. D. Huston, M. Hughes, E. Houpt, and W. A. Petri- Jr. 2003. Current concepts: amoebiasis N Engl J Med 348:1565-1573.

Haque, R., and W. A. Petri- Jr. 2006. Diagnosis of amoebiasis in Bangladesh. Arch Med Res 37:273-276 .

Jittapalapong, S., A. Sangwaranond B. Nimsuphan, T. Inpankaew, C. Phasufk, N. Pinyopanuwat, W. Chimnoi, C. Kengradomkij, P.

Arunwipat, T. Anakewith. 2011. Prevalence of Gastro-Intestinal Parasites of Dairy Cows in Thailand. Kasetsart Journal - Natural Science. 45. 40-45.

Khairnar, K., S.C. Parija. 2007. A novel nested multiplex polymerase chain reaction (PCR) assay for differential detection of Entameoba histolytica, E moshkovskii and E dispar DNA in stool samples BMC Microbiol. 7:47.

Huang, C.C., L.C. Wang, C.H. Pan, C.H.Yang, C.H. Lai. 2014. Investigation of gastrointestinal parasites of dairy cattle around Taiwan; Journal of Microbiology, Immunology and Infection, 47(1) :70-74.

Maharjan, A., J.B. Sherchand, B. Pradhan, A. Paudyal, A.R. Panta. 2007. Rotavirus infection among Diarrheal Children Attending Kanti Children Hospital, Kathmandu, Nepal. Journal of Nepal Health Research Council. 4 : 34-40.

Messmer, T.O., S.K. Skelton, J.F. Moroney, H. Daudharty, B.S. Fields. 2003. Application of a Nested, Multiplex PCR to Psittacosis Outbreaks. Microbiol. 41:5041-5.

Nath, J., S.K. Ghosh, B. Singha, J. Paul. 2015. Molecular Epidemiology of Amoebiasis: A Cross-Sectional Study among North East Indian Population. Haque R, ed. PLoS Neglected Tropical Diseases. 9(12):e0004225.doi:10.1371/journal.pntd.0004225.

Ngui, R., L. Angal, S. H. Fakhrurrazi, Y. L. Ai Lian, L. Y. Ling, J. Ibrahim and R. Mahmud. 2012. Differentiating Entamoeba histolytica, Entamoeba dispar and Entamoeba moshkovskii using nested polymerase chain reaction (PCR) in rural communities in Malaysia, Parasit Vectors. 5: 187.

Parija, S. C., P. K. Prabhakar. 1995. Evaluation of lacto-phenol cotton blue for wet mount preparation of feces J Clin Microbiol. 33:1019–1021.

Pokharel, M., J.B. Sherchand, H.C. Upreti, A. Katuwal, and P. Gauchan. 2007. A Perspective Study on the Etiology of Diarrhea in ChildrenLess than 12 Years of age attending Kanti Children’s Hospital; J. Nepal Paediatr. Soc. 29(1) : 10-16.

Sherchand, J. B. 1997. Intestinal parasitic infection in Southern Nepal. Journal of Institute of Medicine.

Stanley, S. L. Jr. 2003. Amoebiasis Lancet. 361:1025-1034.

Stensvold, C. R., and H.V. Nielsen. 2012. Comparison of Microscopy and PCR for Detection of Intestinal Parasites in Danish Patients Supports an Incentive for Molecular Screening J. Clin. Microbiol. February. 50 (2) : 540-541.

Sykes, A. R. 1994. Parasitism and production in farm animals. Anim. Prod. 59: 155-172.

Timmer, L., C. Visker. 1998. Possibilities and Impossibilities of the use of human excreta as fertiliser in agriculture in sub-Saharan Africa. Royal Tropical Institute and University of Amsterdam, The Netherlands.

Waller, P.J. 1999. International approaches to the concept of integrated control of nematode parasites of livestock. Int. J. Parasitol. 29: 155-164.

WHO. 1997. World Health Organization/Pan American Health Organization/UNESCO report of a consultation of experts on amoebiasis. Wkly Epidemiol Rec WHO. 72: 97-99.




DOI: https://doi.org/10.31186/jspi.id.13.1.101-110

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