Molecular detection and identification of Enteromonas species in human and animal hosts using polymerase chain reaction and DNA sequencing.
Abstract: , a human intestinal protozoan parasite of the diplomonad group, has been overlooked because of its commensal features; therefore, molecular studies on this parasite are limited. To address this gap, we designed a molecular screening protocol using polymerase chain reaction (PCR) and DNA sequencing targeting the 18S small subunit ribosomal RNA gene and applied this screening method to the molecular epidemiological analysis of spp. in humans and various livestock. We validated our methodology using stool samples collected from 215 humans and 270 animal hosts (buffaloes, pigs, dogs, goats, horses, rodents, chickens, and ducks) during an annual epidemiological investigation conducted from 2013 to 2016 on Sumba Island, Indonesia. The overall prevalences of spp. were 33.9 % ( 73/215) in humans and 25.2 % ( 68/270) in mammals and avians. The positive predictive value of this PCR method for spp., as evaluated through sequencing, was 90.1 % in human samples and 58.1 % in non-human samples (particularly low, 11.4 % in rodents). Although the specificity of the PCR approach may not be perfect, in combination with DNA sequencing, it was effective in detecting and identifying a partial sequence (1458 bp) of the target gene region in species.
© 2024 The Author(s).
Publication Date: 2024-07-23 PubMed ID: 39221015PubMed Central: PMC11363483DOI: 10.1016/j.mex.2024.102875Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
Summary
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The research article discusses the use of Polymerase Chain Reaction (PCR) and DNA sequencing in the detection and identification of Enteromonas species, an intestinal parasite found in humans and various livestock. This study provides a detailed analysis of the occurrence of Enteromonas in different hosts and validates the effectiveness of using PCR and DNA sequencing for detection.
Objective of the Study
- The prime focus of the research is to study the Enteromonas species, a type of intestinal parasite that traditionally receives less attention due to its commensal characteristics.
- The researchers aim to design an efficient molecular screening protocol, using Polymerase Chain Reaction (PCR) and DNA sequencing, for the detection and identification of Enteromonas species in various hosts.
Methodology
- The molecular screening protocol targets the 18S small subunit ribosomal RNA gene of the parasite for identification.
- Stool samples were gathered from 215 human and 270 animal subjects, including buffaloes, pigs, dogs, goats, horses, rodents, chickens, and ducks, from Sumba Island, Indonesia, between 2013 and 2016.
Results and Findings
- The overall prevalence of Enteromonas species was found to be 33.9% in humans and 25.2% in mammalian and avian subjects.
- The effectiveness of the PCR method in predicting Enteromonas presence was assessed through DNA sequencing. It was found to be 90.1% accurate in human samples and 58.1% accurate in non-human samples, with especially low accuracy (11.4%) in rodent samples.
Conclusion
- Even though the PCR method alone may not give entirely accurate results, its combination with DNA sequencing proved effective in detecting and identifying the target gene region in Enteromonas species.
Cite This Article
APA
Lacante SA, Jiang C, Mustamir AA, Mizuno T, Bi X, Syafruddin D, Tokoro M.
(2024).
Molecular detection and identification of Enteromonas species in human and animal hosts using polymerase chain reaction and DNA sequencing.
MethodsX, 13, 102875.
https://doi.org/10.1016/j.mex.2024.102875 Publication
Researcher Affiliations
- Department of Global Infectious Diseases, Graduate School of Medical Sciences, Kanazawa University, 13-1 Takaramachi, Kanazawa, Ishikawa, Japan.
- Graduate School of Hasanuddin University, Jl. Perintis Kemerdekaan Km.10, Makassar, Sulawesi Selatan, Indonesia.
- Department of Global Infectious Diseases, Graduate School of Medical Sciences, Kanazawa University, 13-1 Takaramachi, Kanazawa, Ishikawa, Japan.
- Department of Global Infectious Diseases, Graduate School of Medical Sciences, Kanazawa University, 13-1 Takaramachi, Kanazawa, Ishikawa, Japan.
- Graduate School of Hasanuddin University, Jl. Perintis Kemerdekaan Km.10, Makassar, Sulawesi Selatan, Indonesia.
- Department of Global Infectious Diseases, Graduate School of Medical Sciences, Kanazawa University, 13-1 Takaramachi, Kanazawa, Ishikawa, Japan.
- Department of Global Infectious Diseases, Graduate School of Medical Sciences, Kanazawa University, 13-1 Takaramachi, Kanazawa, Ishikawa, Japan.
- Department of Global Infectious Diseases, Advanced Preventive Medical Sciences Research Center, Kanazawa University, 13-1 Takaramachi, Kanazawa, Ishikawa, Japan.
- Department of Parasitology, Faculty of Medicine, Hasanuddin University, Jl. Perintis Kemerdekaan Km.10, Makassar, Sulawesi Selatan, Indonesia.
- Department of Global Infectious Diseases, Graduate School of Medical Sciences, Kanazawa University, 13-1 Takaramachi, Kanazawa, Ishikawa, Japan.
- Department of Global Infectious Diseases, Advanced Preventive Medical Sciences Research Center, Kanazawa University, 13-1 Takaramachi, Kanazawa, Ishikawa, Japan.
Conflict of Interest Statement
The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
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Citations
This article has been cited 1 times.- Jiang C, Lacante SA, Mizuno T, Syafruddin D, Tokoro M. Genetic diversity of genus Chilomastix: molecular classification of C. mesnili and other potential species variations in humans and animals. Trop Med Health 2025 Mar 27;53(1):40.
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