Journal of Advance Researches in Biological Sciences, 2011, 3 (1) 77-79

PREVALENCE OF IGG ANTIBODIES TO AMOEBASIS IN PATIENTS ATTENDING A TERTIARY CARE HOSPITAL IN PUNJAB
Lal Madan1, Berry Vandana2, Swahney Rajesh3, Kaur Rimi4, Singh Gagandeep5 1,2,4,5 Department of Microbiology, Christian Medical College, Ludhiana 3 Department of Applied Biology, Hawassa University, Ethiopia Submitted on:-07/05/2010 Resubmitted on:-07/07/2010

Accepted on:-12/08/2010

Abstract
Amoebiasis is a major cause of morbidity and mortality throughout the tropical world. It can be considered the most aggressive disease of the human intestine responsible in its invasive form for clinical syndromes, ranging from the classical dysentery of acute colitis to extra intestinal disease, with emphasis on hepatic amoebiasis. The study was aimed at determining the seroprevalence of amoebiasis in Ludhiana North India. Nine hundred and twenty three serum samples were tested for IgG antibodies over a period of twelve years from January 1998 to Dec. 2009. Seventy one point eight three percent ( 663/923) of the samples were found to be positive for IgG antibodies. Focus on improved sanitation, personal hygiene, safe disposal of human excreta, protection of water supplies from faecal pollution, avoiding of the use of human excreta as fertilizer, detection and proper treatment of carriers will go a way in prevention and reduction of amoebiasis in the community Key Words- Entamoeba histolytica, Amoebiasis, IgG antibodies.

Introduction
Diarrhoea is a major contributor to mortality and morbidity in the developing world, causing long-term effects on growth and cognitive functions. One of the important etiologies of diarrhoeal diseases is amoebiasis, which is endemic in the developing world. Entamoeba histolytica is an intestinal protozoan parasite and the causative agent of intestinal as well extra intestinal amoebiasis. But the disease remains primarily a problem in the developing world where it leads to 100, 000 deaths per year.1 One of the most puzzling clinical aspects of E. histolytica infection is that 90% of individuals are asymptomatic , whereas remaining 10 % develop colitis, diarrhoea , dysentery and extra intestinal amoebic lesions such as liver abscess. Invasive amoebiasis is an important parasitic disease worldwide in persons residing or traveling to endemic areas.2 It has been estimated that 40 million people develop disabling amoebic colitis or extra intestinal abscess worldwide.3 Amoebic infection is prevalent in the Indian subcontinent, Africa, the Far East and areas of South and Central America. In developing countries it depends largely on cultural habits, level of sanitation, crowding and socio-economic status. Serological examination for anti-amoebic antibodies is a very useful diagnostic study if properly interpreted. After seven days of symptoms, over 95% of patients with amoebic colitis or liver abscess will be seropositive.4

Materials and Methods

Laboratory diagnosis of amoebiasis is usually based on Corresponding Author : Dr. Vandana Berry Department of Microbiology, Christian Medical College, Ludhiana-141002 Punjab, (India) 77

microscopy and serological methods including enzyme linked immunosorbent assay( ELISA), indirect haemagglutination assay ( IAH), and latex agglutination. . Serological tests are more helpful for the identification of E. histolytica infection in industrialized nations, where E. histolytica infection is not common.5 Serum antibodies to E. histolytica can be detected in 75-85 % of patients with symptomatic E. histolytica infection. ELISA is among the most popular methods used in diagnostic laboratories throughout the world. IgG antibody detection was attempted. The study was conducted in the Department of Microbiology, Christian Medical College, Ludhiana, and Punjab. Nine hundred and twenty three serum samples received with clinical suspicion were tested serologically for the detection of IgG antibodies against E .histolytica specific lectin coproantigen using Ridascreen kit (R Biopharm AG Landwehrstr-540 DARMSTADT, Germany). Serum samples were diluted 1:50 with buffer . 100 ul of diluted serum was placed into the well and incubated at 25 0C for 15 minutes. Positive and negative controls were run in parallel. After washing for 5 times, 100ul of conjugate was added to each well and incubated at 25 0 C for 15 minutes. Well were washed again as per the manufacturer's advice. 50 micolit of substrate and 50 ul of chromogen was added to each well. Wells were placed at room temperature for 15 minutes. Reaction was stopped by adding a drop of stop solution after 15 minutes. Samples were photometrically measured at 450 nm as per the manufacturers' advice. Samples giving the Index value of more than 1.1 were considered to be positive while Index value less than 1.1 was taken as negative. Data was stastically analysed for linear trends using chi square, p value was 0,0362 while the chi square for linear trend showed value of

Journal of Advance Researches in Biological Sciences, 2011, 3 (1) 77-79

4.514 indicating the difference of proportions in this series is marginally significant. abscess patients9. A study from Bangladesh revealed a prevalence of 52% in children that were colonized with E.histolotica10. Seroepidemiologic investigations of amebiasis in some tropical areas of Mexico indicate that while the prevalence of anti-amebic antibodies is relatively low in areas where epidemic transmission has not been reported, during epidemics an incidence rate of 50% is common, reaching as high as 80% during epidemics11. E.histolitica appears to be more virulent than other opportunistic pathogens as it can cause severe infections in both immunocompetent and immunocompromised hosts 4. There is a 71.83% prevalence of amoebiasis estimated in Microbiology Laboratory of CMC and Hospital, Ludhiana, a tertiary care hospital. Hence the figure is reflecting high prevalence rate of amoebiasis not only in Ludhiana but also in the adjoining regions of Punjab as well as of the other states from where patients come to CMC for treatment. This distressing disease, associated with numerous complications is basically a disease of the developing nations with improper means of sanitation. Health education of public regarding personal cleanliness and ingestion of safe food and water can help a lot in reducing the prevalence of this infection. In the community there is a great need to focus on effective sanitation, safe disposal of human excreta, protection of water supplies from faecal pollution and avoiding the use of human excreta as a fertilizer. All the carriers in the community need to be traced and treated properly. Moreover the amount spent on the prophylaxis of amoebiasis is definitely meager than that spent on treating manifestations incurred by this disease.

Results and Discussion

Nine hundred and twenty three samples were tested over a period of 12 years from January 1998 to Dec.2009 for IgG antibodies against E. histolytica. Seventy one point eight three percent (663/923) of the samples were found to be positive.. Year wise results are shown in the table 1. The prevalence is running between 57.14% and 87.93%. The rate remains significant over the years with fluctuating pattern. There is some fall in the degree of prevalence in the recent years i.e., 2008 and 2009. Geographically no clustering was seen in the cases as Christian Medical College is a tertiary care hospital and patients come to CMC from neibouring states of Haryana, Rajasthan, Himachal, Uttaranchal and Uttar Pradesh. Eighty percent of the patients were adults and 78% males. E. histolytica remains a significant cause of morbidity and mortality in developing countries6. E.histolytica usually causes asymptomatic infections but in a minority of cases causes symptoms ranging from a few loose stools to bloody diarrhoea 7 .infection. Extra intestinal manifestations in amoebiasis include amoebic liver abscess, pulmonary amoebiasis, cerebral amoebiasis, splenic abscess and cutaneous lesions. Various studies done in India report varying prevalence in different regions.(sentence framing) A study from Patna, Bihar reported much lower rate of 27% antibodies development in E.histolytica infected subjects8. Another study from New Delhi reported comparable results of 79% from Amoebic liver Table 1: Amoebic Serology Depicting IgG Antibodies. Year 1998 1999 2000 2001 2002 2003 2004 2005 2006 2007 2008 2009 Total Total No. of Sample 56 26 60 62 64 81 129 116 103 95 70 61 923

Positive 42 17 47 43 48 57 92 102 73 67 40 35 663

& 74.99 65.38 78.33 69.35 75.00 70.37 71.31 87.93 70.87 70.52 57.14 57.35 71.83

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Journal of Advance Researches in Biological Sciences, 2011, 3 (1) 77-79

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