scholarly journals A Comparative Analysis of Microscopy, Coproantigen Serology, and Nested Multiplex PCR in the Laboratory Diagnosis of Entamoeba histolytica Infection

2021 ◽  
Author(s):  
Sindhusuta Das ◽  
Nonika Rajkumari ◽  
Anitha Gunalan ◽  
Dhanalakshmi Rajavelu ◽  
Jeby Jose Olickal
Keyword(s):  
Multiplex Pcr ◽  
Entamoeba Histolytica ◽  
Predictive Value ◽  
Parasitic Infection ◽  
Laboratory Diagnosis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Kappa Statistics ◽  
Histolytica Infection ◽  
Intestinal Amoebiasis ◽  
Stool Samples

Abstract Objectives Amoebiasis is caused by the most common intestinal protozoan parasite Entamoeba histolytica. This parasite causes amoebic colitis, which is manifested by diarrhea, followed by dysentery. The laboratory diagnosis of intestinal amoebiasis in most cases is by microscopic examination of stool samples. Other nonroutine methods include coproantigen enzyme-linked immunosorbent assay (ELISA) from stool samples, serum ELISA for antibodies, stool culture, isoenzyme analysis, and polymerase chain reaction (PCR). The present study aimed to comparatively analyze the different diagnostic modalities used for the detection of E. histolytica from the stool sample of patients with intestinal amoebiasis. Materials and Methods This study was undertaken with 631 patients, during a period of 3 years, from January 2017 to December 2019. Stool specimen obtained from each patient was subjected to direct microscopic wet mount examination, coproantigen ELISA, and nested multiplex PCR, respectively. Results Out of all the patients tested, 5.2% were positive for E. histolytica. Among the positive cases, stool microscopy was positive in 3.17%, coproantigen ELISA was positive in 29 (4.6%) cases, and PCR was positive in 30 (4.75%) cases. Statistical Analysis The prevalence of E. histolytica infection was summarized as percentages. The three diagnostic tests done were statistically analyzed, taking microscopy as the gold standard. The agreement between techniques (microscopy, coproantigen ELISA, and PCR) was analyzed with kappa statistics. Sensitivity, specificity, positive predictive value, negative predictive value, and diagnostic accuracy were summarized as percentage with 95% confidence interval. Conclusion In all suspected amoebiasis cases, a combination of stool microscopy, coproantigen testing with molecular detection of the parasite offers the best approach to diagnosis of this parasitic infection.

scholarly journals High occurrence of Entamoeba histolytica in the municipalities of Ariquemes and Monte Negro, State of Rondônia, Western Amazonia, Brazil

2013 ◽  
Vol 55 (3) ◽  
pp. 193-196 ◽  
Author(s):  
Rafael Vital dos Santos ◽  
Jucélia da Silva Nunes ◽  
Juliana Almeida de Souza Aranha Camargo ◽  
Eliana Maria Maurício da Rocha ◽  
Gilberto Fontes ◽  
...  
Keyword(s):  
Entamoeba Histolytica ◽  
Odds Ratio ◽  
Detection Rate ◽  
Enzyme Linked Immunosorbent Assay ◽  
High Occurrence ◽  
Significant Difference ◽  
Histolytica Infection ◽  
Stool Antigen ◽  
Stool Samples ◽  
Two Populations

Introduction: Entamoeba histolytica infections were investigated in residents of the Ariquemes and Monte Negro municipalities in Rondônia State, Brazil. Methods: Stool samples of 216 individuals were processed by the spontaneous sedimentation method and analyzed by microscopy for detection of the E. histolytica/E. dispar complex, followed by the immunoassay method using an enzyme-linked immunosorbent assay-based kit for the E. histolytica stool antigen. Results: E. histolytica/E. dispar cysts were present in 61% (50/82) and 44% (59/134) of the samples from Ariquemes and Monte Negro respectively, with a significant difference in the occurrence of infection between the two populations [p < 0.05; χ2 = 5.2; odds ratio = 2.0 (1.1 - 3.6)]. The E. histolytica antigen detection rate was 36.6% (30/82) for stool samples from Ariquemes, and 19.4% (26/134) for stool taken from the residents of Monte Negro. The rate of the occurrence of amoebiasis was significantly higher in the population from Ariquemes [p < 0.05; χ2 = 7.8; odds ratio = 2.4 (1.2 - 4.7)]. Discussion: Due to the high occurrence of E. histolytica infected residents diagnosed in the region and the unavailability in local clinics of a test to distinguish between the two Entamoeba species, physicians should consider treating E. histolytica/E.dispar infections. Conclusion: The results indicate that E. histolytica infection is highly endemic in the studied areas.

scholarly journals Detection of Entamoeba histolytica Coproantigen Among Children with Dysentery in Ahvaz, Southwest Iran

2020 ◽  
Vol In Press (In Press) ◽  
Author(s):  
Ahmad Shamsizadeh ◽  
Roya Nikfar ◽  
Mahmoud Rahdar
Keyword(s):  
Entamoeba Histolytica ◽  
False Positive ◽  
Correct Diagnosis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Medical Laboratories ◽  
Negative Results ◽  
Histolytica Infection ◽  
Stool Samples ◽  
Southwest Iran ◽  
Diagnosis Of Infection

: Entamoeba histolytica is one of the important parasitic diseases in many parts of the world, especially the tropical and subtropical regions. The parasite is transmitted through contaminated water and vegetables. The exact diagnosis of infection with the parasite is crucial in many medical laboratories since there are many false positive and negative results in their reports. Therefore, the current study aimed at evaluating and comparing microscopic and coproantigen ELISA (the enzyme-linked immunosorbent assay) results to reach an appropriate test for the correct diagnosis of amoebiasis in children. One hundred stool samples were collected from children under 15 years old with dysenteric diarrhea from April to September 2018. Microscopic tests and coproantigen ELISA were performed on all the samples. The results showed that 5% of the samples had E. histolytica/E. dispar cysts. The findings of ELISA to detect coproantigen did not show any specific E. histolytica antigen in the samples. Hence, all the patients received chemotherapy for shigellosis. E. histolytica infection is not the main causative agent for dysenteric diarrhea in children in the studied area, and laboratory experts should be trained to prevent false-positive reports.

scholarly journals The NF-κB p50 Subunit Is Protective during Intestinal Entamoeba histolytica Infection of 129 and C57BL/6 Mice

2010 ◽  
Vol 78 (4) ◽  
pp. 1475-1481 ◽  
Author(s):  
Kyou-Nam Cho ◽  
Stephen M. Becker ◽  
Eric R. Houpt
Keyword(s):  
Entamoeba Histolytica ◽  
Electrical Resistance ◽  
Transepithelial Electrical Resistance ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mobility Shift ◽  
Protective Function ◽  
Targeted Deletion ◽  
Nuclear Extracts ◽  
Histolytica Infection ◽  
Mobility Shift Assay

ABSTRACT Entamoeba histolytica is the agent of amebic colitis. In this work we examined the intestinal NF-κB response to this parasite. Using an enzyme-linked immunosorbent assay (ELISA) and an electrophoretic mobility shift assay, we found that the NF-κB subunit p50 predominated in nuclear extracts of whole cecal tissue and of isolated crypts from mice inoculated with E. histolytica. p50 was protective, since C57BL/6 and 129 mice in which there was targeted deletion of this subunit were more susceptible to E. histolytica infection as measured by culture results, cecal parasite ELISA results, and/or histologic scores. The transepithelial electrical resistance of cecal explants from C57BL/6 and 129 p50 knockout mice decreased markedly in response to the parasite compared with the transepithelial electrical resistance of their wild-type counterparts, suggesting that a protective function of p50 was present in the epithelium itself. This work shows that NF-κB activity, particularly activity of the p50 subunit, is one factor that contributes to resistance of the gut to E. histolytica infection.

scholarly journals Intestinal Parasite Infestation in HIV Infected Patients in Tertiary Care Center

2021 ◽  
Vol 15 (3) ◽  
pp. 1602-1607
Author(s):  
Dhruba Hari Chandi ◽  
Sucheta J. Lakhani
Keyword(s):  
Entamoeba Histolytica ◽  
Cryptosporidium Parvum ◽  
Intestinal Parasite ◽  
Intestinal Parasites ◽  
Parasitic Infection ◽  
Intestinal Parasitic Infection ◽  
Coccidian Parasites ◽  
Stool Samples ◽  
Taenia Species ◽  
Hiv Seropositive

Every year, the number of people living with HIV rises as a consequence of advanced infections and the positive effects of highly active antiretroviral therapy (HAART). Gastrointestinal involvement is common, with 90% of patients seeking treatment for gastrointestinal problems as their HIV infection progresses. Nonetheless, identifying and characterization of infectious agents is important for patient management by excluding a clinical diagnosis and determining appropriate treatment, as well as determining public healthcare policy for true pathogen prevalence and yielding epidemiological risk factors for specific infections. The aim of this study is to evaluate the prevalence of symptomatic or asymptomatic intestinal parasitic infection among HIV or AIDS patients. For this study with 80 HIV seropositive patients being recruited from various wards and the Integrated Counseling and Testing Center (ICTC) affiliated to the microbiology department. Patients with acute and chronic diarrhea with abdominal disorder were taken as symptomatic whereas patients without these clinical complaints and who came for routine investigations were taken as asymptomatic. Firstly stool samples were analyzed by macroscopically for the presence of mucus, blood, larvae, segments of tapeworm and adult worms. The consistencies of stool were also recorded such as formed, watery or soft or loose with odor and color. It was examined microscopically after macroscopically for protozoan cysts and trophozoites, helminthic ova and larvae, as wet mount preparation by saline and iodine preparation as well as formal ether concentrated. For the detection of intestinal coccidian parasites, smears were prepared from stool samples and a modified Ziel-Nelseen (MZN) stain was also performed. Stool samples with the detection of parasites were informed for treatment. The prevalence of intestinal parasite was 23.75% with asymptomatic and symptomatic groups having a prevalence of 16.98% and 37.04% respectively. Out of total patients, 56.25% were male and 43.75% were female. Among the male patients, 11 (13.75%) were positive for an intestinal parasitic infection and 8 (10%) were positive among females. The age distribution data revealed that the age group 21-40 years old had the highest number of intestinal parasites, followed by 41-60, 61-80, and 0-20 years old. The most intestinal parasites were found in the young and middle-aged patients, according to this study. Different parasites were identified as Entamoeba histolytica, Taenia species, Ascaris lumbricoides, Cryptosporidium parvum and Isospora with one protozoan, two coccidian parasites and two helminthes. The most common parasite was Taenia species 6 (7.5%) followed by Entamoeba histolytica 5 (6.3%) and Cryptosporidium parvum 5 (6.3%). Intestinal parasitic infection is not uncommon in HIV seropositive patients. This study underscores the need for early diagnosis and treatment of these intestinal parasites in both symptomatic and asymptomatic HIV patients.

scholarly journals Prevalence and associated risk factors of Entamoeba histolytica, E. dispar and E. moshkovskii infection among Orang Asli communities in Slim River, Perak

2020 ◽  
Vol 8 (2) ◽  
pp. 22-35
Author(s):  
Syazwan Saidin ◽  
Adibah Abu Bakar ◽  
Badrul Munir Mohd Zain
Keyword(s):  
Risk Factors ◽  
Rural Area ◽  
Entamoeba Histolytica ◽  
Gastrointestinal Symptom ◽  
Pcr Analysis ◽  
Orang Asli ◽  
Hygiene Practices ◽  
Histolytica Infection ◽  
Stool Samples ◽  
Associated Risk Factors

Entamoeba infection is still widespread in Malaysia’s rural area particularly among Orang Asli communities which usually associated with poverty and lack of sanitation. Due to scarce information on these Entamoeba infections in Slim River, Perak we conducted this study to investigate the prevalence and associated risk factors towards this infection. A total of 55 stool samples from voluntary participants with and without symptoms of amoebiasis were collected and examined using PCR technique. PCR analysis showed 16.4% stool samples were detected positive for Entamoeba spp., discriminated as 7.3% that were positive for E. histolytica and 9.1% for E. dispar. No E. moshkovskii was detected at all. Factors such as indiscriminate defecation, improper sewage disposal and not washing hand after playing with soil or gardening showed significant association with E. histolytica infection; while gastrointestinal symptom such as vomiting was associated with E. dispar infection. In conclusion, the study reveals there is still an occurrence of Entamoeba spp. among Orang Asli communities in Slim River, Perak. This implies that good personal and hygiene practices should be enhanced through awareness strategy to control Entamoeba infections in Orang Asli communities in Malaysia.

scholarly journals Clostridium difficile PCR Cycle Threshold Predicts Free Toxin

2017 ◽  
Vol 55 (9) ◽  
pp. 2651-2660 ◽  
Author(s):  
Fiona Senchyna ◽  
Rajiv L. Gaur ◽  
Saurabh Gombar ◽  
Cynthia Y. Truong ◽  
Lee F. Schroeder ◽  
...  
Keyword(s):  
Clostridium Difficile ◽  
Predictive Value ◽  
Reference Method ◽  
Neutralization Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Content Type ◽  
Cycle Threshold ◽  
Positive Stool ◽  
Stool Samples ◽  
Sensitivity Specificity

ABSTRACTThere is no stand-aloneClostridium difficilediagnostic that can sensitively and rapidly detect fecal free toxins. We investigated the performance of theC. difficilePCR cycle threshold (CT) for predicting free toxin status. Consecutive stool samples (n= 312) positive for toxigenicC. difficileby the GeneXpertC. difficile/EpitcdBPCR assay were tested with the rapid membrane C. Diff Quik Chek Complete immunoassay (RMEIA). RMEIA toxin-negative samples were tested with the cell cytotoxicity neutralization assay (CCNA) and tgcBIOMICS enzyme-linked immunosorbent assay (ELISA). Using RMEIA alone or in combination with CCNA and/or ELISA as the reference method, the accuracy ofCTwas measured at differentCTcutoffs. Using RMEIA as the reference method, aCTcutoff of 26.35 detected toxin-positive samples with a sensitivity, specificity, positive predictive value, and negative predictive value of 96.0% (95% confidence interval [CI], 90.2% to 98.9%), 65.9% (95% CI, 59.0% to 72.2%), 57.4% (95% CI, 52.7% to 62%), and 97.1% (95% CI, 92.8% to 98.9), respectively. Inclusion of CCNA in the reference method improvedCTspecificity to 78.0% (95% CI, 70.7% to 84.2%). Intercartridge lotCTvariability measured as the average coefficient of variation was 2.8% (95% CI, 1.2% to 3.2%). Standardizing the input stool volume did not improveCTtoxin specificity. The medianCTvalues were not significantly different between stool samples with Bristol scores of 5, 6, and 7, between pediatric and adult samples, or between presumptive 027 and non-027 strains. In addition to sensitively detecting toxigenicC. difficilein stool, on-demand PCR may also be used to accurately predict toxin-negative stool samples, thus providing additional results in PCR-positive stool samples to guide therapy.

scholarly journals Parasitic infections and risk factors associated with Amoebiasis among pregnant women attending antenatal clinics in primary health care centres in Lagos Mainland, Lagos, Nigeria

2019 ◽  
Vol 1 ◽  
pp. 52-60
Author(s):  
B Akinsanya ◽  
A Babatunde ◽  
M Olasanmi ◽  
A A Adedotun
Keyword(s):  
Risk Factors ◽  
Health Care ◽  
Primary Health Care ◽  
Pregnant Women ◽  
Entamoeba Histolytica ◽  
Mixed Infection ◽  
Ascaris Lumbricoides ◽  
Primary Health ◽  
Histolytica Infection ◽  
Stool Samples

The prevalence of amoebiasis was investigated among pregnant women using a gold standard microscopy method. Stool samples were collected from pregnant women, from two primary health care centers (Iwaya and Ebute-metta). The study involved 203 pregnant women during their visits to the antenatal clinic in Primary health centres at Lagos Mainland from June – October, 2016. Each participant was interviewed using questionnaires to relate prevalence and risk factors to their socio-demographic characteristics. Among these pregnant women, 120 respondents provided their stool samples and this was parasitologically screened with 0.8% prevalence of Entamoeba histolytica infection, 7.5% with Entamoeba Coli, 0.5% with Ascaris lumbricoides, and 0.8% infection with Hookworm. A prevalence of 0.8% for the mixed infection of Ascaris lumbricoides with Entamoeba histolytica, and a prevalence of 1.7% for the mixed infection of Entamoeba coli and Entamoeba histolytica was recorded. Significant association (p<0.05), was established between infection and diarrhoea, and the residence of the pregnantwomen.Socio - demographic factors like age, sex, socio-economic status, type of toilet, source of drinking water (p>0.05) showed no significant association with amoebiasis prevalence. In conclusion, E. histolytica infection is generally low in Lagos state owing to good environmental sanitation within Lagos metropolis. Efforts should therefore be geared towards its elimination by provision of water for Lagos residents.

scholarly journals Multisite Clinical Evaluation of a Rapid Test for Entamoeba histolytica in Stool

2014 ◽  
Vol 53 (2) ◽  
pp. 493-497 ◽  
Author(s):  
Hans P. Verkerke ◽  
Blake Hanbury ◽  
Abdullah Siddique ◽  
Amidou Samie ◽  
Rashidul Haque ◽  
...  
Keyword(s):  
Entamoeba Histolytica ◽  
Point Of Care ◽  
Rapid Test ◽  
Independent Study ◽  
Enzyme Linked Immunosorbent Assay ◽  
Microplate Assay ◽  
Content Type ◽  
Stool Samples ◽  
Point Of Care Detection ◽  
Enteric Protozoa

Rapid point-of-care detection of enteric protozoa in diarrheal stool is desirable in clinical and research settings to efficiently determine the etiology of diarrhea. We analyzed the ability of the third-generationE. histolyticaQuik Chek assay developed by Techlab to detect amebic antigens in fecal samples collected from independent study populations in South Africa and Bangladesh. We compared the performance of this recently released rapid test to that of the commercially available ProSpecTEntamoeba histolyticamicroplate assay from Remel and theE. histolyticaII enzyme-linked immunosorbent assay (ELISA) from Techlab, using real-time and nested-PCR forEntamoebaspecies to resolve any discrepant results. After discrepant resolution, TheE. histolyticaQuik Chek assay exhibited sensitivity and specificity compared to theE. histolyticaII ELISA of 98.0% (95% confidence interval [CI], 92.9% to 99.8%) and 100% (95% CI, 99.0% to 100%), respectively. Compared to the ProSpecT microplate assay, theE. histolyticaQuik Chek (Quik Chek) assay exhibited 97.0% sensitivity (95% CI, 91.5% to 99.4%) and 100% specificity (95% CI, 99.0% to 100%). Our results indicate that the Quik Chek is a robust assay for the specific detection ofE. histolyticatrophozoites in unfixed frozen clinical stool samples.

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