Laboratory methods of identification of Entamoeba histolytica and its differentiation from look-alike Entamoeba spp.

Parija, Subhash Chandra; Mandal, Jharna; Ponnambath, Dinoop Korol

doi:10.4103/2229-5070.138535

Cited by 46 publications

(43 citation statements)

References 40 publications

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“…Even the commercial ELISA-based method for specific identification and detection of E. histolytica in fecal specimens [23] has shown poor sensitivity and specificity in many studies because of cross-contaminations with other parasites [24,25] . In the last decade, molecular-based diagnostic tests have gained importance in the diagnosis of many infectious diseases including amoebiasis to overcome the problems of conventional methods with the advantages of increased sensitivity, specificity, and simplicity [26,27] . Several PCR assays designed to differentiate E. histolytica from E. dispar have been described [15,28,29] .…”

Section: Discussionmentioning

confidence: 99%

Differentiation of Entamoeba histolytica from Entamoeba dispar by nested multiplex polymerase chain reaction

Mohammed

Taha

2017

Parasitologists United Journal

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Background: Amoebiasis is a parasitic disease caused by the intestinal protozoon Entamoeba histolytica. Microscopic examination fails to differentiate E. histolytica from the morphologically identical nonpathogenic Entamoeba dispar. To avoid unnecessary treatment of individuals infected with nonpathogenic E. dispar, it is essential to differentially diagnose infections caused by pathogenic from nonpathogenic Entamoeba spp. Objective: The aim of this study was to assess the efficacy of nested multiplex PCR (NM PCR) technique as a diagnostic method for differentiating infections caused by E. histolytica and E. dispar. Materials and methods: Stool samples collected from patients with and without symptoms of amoebiasis were screened for E. histolytica/E. dispar trophozoites/cysts by microscopic examination. NM PCR was performed on a total of 52 samples targeting the genus-specific 16S-like ribosomal RNA gene for simultaneous, differential detection of E. histolytica and E. dispar. Results: NM PCR detected E. histolytica at 439 bp and E. dispar at 174 bp, and it was positive in 31 out of 32 cases with a sensitivity of 96.85%. From those, 17 (32.7%) samples were positive for E. histolytica, 12 (23.1%) for E. dispar, and three (5.7%) for both species. In addition, NM PCR diagnosed E. dispar in one of the negative controls with a specificity of 95%. Conclusions: NM PCR is useful for the specific detection of E. histolytica and E. dispar in stool samples. Additional methods for species differentiation have been used to avoid unnecessary treatment of individuals infected with the nonpathogenic species [12]. PCR demonstrated excellent sensitivity and specificity

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Section: Discussionmentioning

confidence: 99%

Differentiation of Entamoeba histolytica from Entamoeba dispar by nested multiplex polymerase chain reaction

Mohammed

Taha

2017

Parasitologists United Journal

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“…Correct data on the pathogen load of E. histolytica in stool samples of patients are not readily available, as the evaluation by microscopy in stool is unreliable and cannot discern between pathogenic and apathogenic Entamoeba spp. [14]. Older literature data suggest that Entamoeba infection will be detected by microscopy in two stool examinations at a cyst excretion rate of 100.000 to 1.000.000 cysts per day [15].…”

Section: Methodsmentioning

confidence: 99%

Detection of Gastrointestinal Pathogens from Stool Samples on Hemoccult Cards by Multiplex PCR

Alberer

Schlenker

Bauer

et al. 2017

Canadian Journal of Infectious Diseases and Medical Microbiolog

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Purpose. Up to 30% of international travelers are affected by travelers' diarrhea (TD). Reliable data on the etiology of TD is lacking. Sufficient laboratory capacity at travel destinations is often unavailable and transporting conventional stool samples to the home country is inconvenient. We evaluated the use of Hemoccult cards for stool sampling combined with a multiplex PCR for the detection of model viral, bacterial, and protozoal TD pathogens. Methods. Following the creation of serial dilutions for each model pathogen, last positive dilution steps (LPDs) and thereof calculated last positive sample concentrations (LPCs) were compared between conventional stool samples and card samples. Furthermore, card samples were tested after a prolonged time interval simulating storage during a travel duration of up to 6 weeks. Results. The LPDs/LPCs were comparable to testing of conventional stool samples. After storage on Hemoccult cards, the recovery rate was 97.6% for C. jejuni, 100% for E. histolytica, 97.6% for norovirus GI, and 100% for GII. Detection of expected pathogens was possible at weekly intervals up to 42 days. Conclusion. Stool samples on Hemoccult cards stored at room temperature can be used in combination with a multiplex PCR as a reliable tool for testing of TD pathogens.

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“…They are all morphologically indistinguishable. Serologic tests have been developed, which are less sensitive than PCR for 16 S rDNA (68, 93). The diagnosis of large tapeworms is similarly difficult primarily due to a newly identified species Taeina asiatica .…”

Section: Application Of Molecular Techniques To the Epidemiology Of Ementioning

confidence: 99%

Population Genetics and Molecular Epidemiology of Eukaryotes

Blanton

2018

Microbiol Spectr

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SUMMARY: Molecular epidemiology uses the distribution and organization of a pathogen’s DNA to understand the distribution and determinants of disease. Since the biology of DNA for eukaryotic pathogens differs substantially from that of bacteria, the analytic approach to their molecular epidemiology can also differ. While many of the genotyping techniques presented earlier in this series, Advances in Molecular Epidemiology of Infectious Diseases, can be applied to eukaryotes, the output must be interpreted in the light of how DNA is distributed from one generation to the next. In some cases, parasite populations can be evaluated in ways reminiscent of bacteria. They differ, however, when analyzed as sexually reproducing organisms, where all individuals are unique, but the genetic composition of the population does not change unless a limited set of events occurs. It is these events (migration, mutation, non-random mating, selection, genetic drift) that are of interest. At a given time, not all of them are likely to be equally important, so the list can easily be narrowed down to understand the driving forces behind the population as it is now and even what it will look like in the future. The main population characteristics measured to assess these events are differentiation and diversity, interpreted in the light of what is known about the population from observation. The population genetics of eukaryotes is important for planning and evaluation of control measures, surveillance, outbreak investigation, and monitoring the development and spread of drug resistance.

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Laboratory methods of identification of Entamoeba histolytica and its differentiation from look-alike Entamoeba spp.

Cited by 46 publications

References 40 publications

Differentiation of Entamoeba histolytica from Entamoeba dispar by nested multiplex polymerase chain reaction

Differentiation of Entamoeba histolytica from Entamoeba dispar by nested multiplex polymerase chain reaction

Detection of Gastrointestinal Pathogens from Stool Samples on Hemoccult Cards by Multiplex PCR

Population Genetics and Molecular Epidemiology of Eukaryotes

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