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Diagnosis of amphimeriasis by LAMPhimerus assay in human stool samples long-term storage onto filter paper
- Source :
- PLoS ONE, Vol 13, Iss 2, p e0192637 (2018), PLoS ONE
- Publication Year :
- 2018
- Publisher :
- Public Library of Science (PLoS), 2018.
-
Abstract
- Amphimeriasis, a fish-borne zoonotic disease caused by the liver fluke Amphimerus spp., has recently been reported as an emerging disease affecting an indigenous Ameridian group, the Chachi, living in Ecuador. The only method for diagnosing amphimeriasis was the microscopic detection of eggs from the parasite in patients' stool samples with very low sensitivity. Our group developed an ELISA technique for detection of anti-Amphimerus IgG in human sera and a molecular method based on LAMP technology (named LAMPhimerus) for specific and sensitive parasite DNA detection. The LAMPhimerus method showed to be much more sensitive than classical parasitological methods for amphimeriasis diagnosis using human stool samples for analysis. The objective of this work is to demonstrate the feasibility of using dried stool samples on filter paper as source of DNA in combination with the effectiveness of our previously designed LAMPhimerus assay for successfully Amphimerus sp. detection in clinical stool samples. A total of 102 untreated and undiluted stool samples collected from Chachi population were spread as thin layer onto common filter paper for easily transportation to our laboratory and stored at room temperature for one year until DNA extraction. When LAMPhimerus method was applied for Amphimerus sp. DNA detection, a higher number of positive results was detected (61/102; 59.80%) in comparison to parasitological methods (38/102; 37.25%), including 28/61 (45.90%) microscopy-confirmed Amphimerus sp. infections. The diagnostic parameters for the sensitivity and specificity werecalculated for our LAMPhimerus assay, which were 79.17% and 65.98%, respectively. We demonstrate, for the first time, that common filter paper is useful for easy collection and long-term storage of human stool samples for later DNA extraction and molecular analysis of human-parasitic trematode eggs. This simple, economic and easily handling method combined with the specific and sensible LAMPhimerus assay has the potential to beused as an effective molecular large-scale screening test for amphimeriasis-endemic areas.
- Subjects :
- Male
0301 basic medicine
Veterinary medicine
Molecular biology
lcsh:Medicine
Molecular biology assays and analysis techniques
Biochemistry
Zoonotic disease
Feces
0302 clinical medicine
Filter Paper
Specimen Storage
Nucleic Acids
Zoonoses
Medicine and Health Sciences
Parasite hosting
Child
lcsh:Science
DNA extraction
education.field_of_study
Multidisciplinary
Eukaryota
Opisthorchidae
Middle Aged
Liver fluke
Molecular analysis
Laboratory Equipment
Freshwater Fish
Child, Preschool
Vertebrates
Engineering and Technology
Female
Research Article
Adult
Adolescent
030231 tropical medicine
030106 microbiology
Population
Equipment
Enzyme-Linked Immunosorbent Assay
Biology
Young Adult
03 medical and health sciences
Extraction techniques
Parasitic Diseases
Genetics
Animals
Humans
In patient
DNA filter assay
education
Aged
Filter paper
lcsh:R
Organisms
Biology and Life Sciences
Infant
DNA
Research and analysis methods
Molecular biology techniques
Fish
Storage and Handling
Parasitology
lcsh:Q
Subjects
Details
- Language :
- English
- ISSN :
- 19326203
- Volume :
- 13
- Issue :
- 2
- Database :
- OpenAIRE
- Journal :
- PLoS ONE
- Accession number :
- edsair.doi.dedup.....2d402c24025550e93eb0397508c4a71d