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Open veterinary journal2024; 14(4); 1051-1058; doi: 10.5455/OVJ.2024.v14.i4.12

Detection of the chuA gene encoding the invasive enterohemorrhagic species Escherichia coli 0157:H7 using qPCR in horse feces samples on Sumbawa Island, Indonesia.

Abstract: Bacterial identification can be done using various testing techniques. Molecular techniques are often used to research dangerous diseases, an approach using genetic information on the pathogenic agent. The enterohemorrhagic invasive species 0157:H7 was identified from the feces of working horses on the island of Sumbawa. Another advance in molecular technology is genome amplification with qPCR which is the gold standard for detecting Unassigned: This study aims to detect and identify the invasive species 0157:H7 using the gene encoding with the qPCR method sourced from horse feces. Unassigned: Fresh fecal samples from horses on Sumbawa Island were isolated and identified, then continued with molecular examination using the gene encoding using the qPCR method. Unassigned: qPCR testing in this study showed that six sample isolates that were positive for 0157:H7 were detected for the presence of the gene, which is a gene coding for an invasive species of bacteria. The highest to lowest Cq values and Tm from the qPCR results of the sample isolates were 15.98 (4KJ), 14.90 (19KG), 14.6 (3KJ), 13.77 (20KG), 12.56 (5KGB), and 12.20 (6KJ). Tm values are 86.7 (4KJ), 86.69 (3KJ), 86.56 (5KGB), 85.88 (20KGB), 85.81 (19KG), and 85.74 (6KJ). Unassigned: Validation, standardization of the development, and modification of qPCR technology must be carried out to harmonize testing throughout to avoid wrong interpretation of the test results so that the determination of actions to eradicate and control diseases originating from animals in the field does not occur.
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Publication Date: 2024-04-30 PubMed ID: 38808295PubMed Central: PMC11128647DOI: 10.5455/OVJ.2024.v14.i4.12Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

Detection of the chuA gene associated with the invasive enterohemorrhagic Escherichia coli O157:H7 was successfully carried out on horse feces from Sumbawa Island, Indonesia, using quantitative PCR (qPCR) methods, indicating the presence of this pathogenic bacterium.

Background and Objectives

  • Escherichia coli O157:H7 is a dangerous enterohemorrhagic invasive bacterial species that can cause serious diseases.
  • Molecular identification techniques, particularly genetic-based methods, are crucial for detecting pathogenic bacteria in samples.
  • The study aimed to detect the chuA gene, which encodes for the invasive O157:H7 species, using quantitative PCR (qPCR).
  • Horse fecal samples from Sumbawa Island, Indonesia, were chosen as the source to assess the presence of this pathogen in a working animal population.

Methodology

  • Fresh fecal samples were collected from working horses on Sumbawa Island.
  • Bacterial isolates were obtained from these samples using standard microbiological isolation techniques.
  • The isolates then underwent molecular analysis via qPCR to detect the presence of the chuA gene.
  • Quantitative PCR was used because it is a sensitive and rapid technique and considered a gold standard for detecting specific genes related to pathogens.

Results

  • Six of the isolated samples tested positive for the chuA gene, confirming the presence of E. coli O157:H7 in horse feces.
  • The samples showed a range of Cq (cycle quantification) values from 12.20 to 15.98, indicating varying levels of bacterial DNA quantity among the isolates.
    • Higher Cq values correspond to lower initial quantities of target DNA, and lower Cq values indicate higher bacterial load.
  • Melting temperature (Tm) analysis of the qPCR products ranged between 85.74°C and 86.7°C, providing confirmation of specific amplification of the targeted gene.
  • The consistency of Tm among isolates supports the specificity of the qPCR assay for the chuA gene detection.

Interpretation and Significance

  • Detection of the chuA gene in horse feces highlights a potential reservoir and source of E. coli O157:H7 that may pose health risks to humans and animals.
  • These findings demonstrate the utility of qPCR for rapid and accurate detection of invasive pathogen genes in environmental or animal samples.
  • The study underscores the importance of validating and standardizing qPCR protocols to ensure reproducibility and avoid misinterpretation of results.
  • Reliable molecular detection supports effective disease control and eradication strategies by identifying pathogenic bacteria in the field.
  • Monitoring pathogens in working animals like horses can help control zoonotic transmission routes and safeguard public health.

Recommendations

  • Further development, validation, and harmonization of qPCR assays are required for consistent field and laboratory testing outcomes.
  • Continuous surveillance of bacterial pathogens in animal populations is important for early detection and response to potential outbreaks.
  • Implementation of control measures based on accurate molecular diagnostics can reduce disease transmission from animals to humans.

Cite This Article

APA
Kholik K, Sukri A, Riwu KHP, Kurniawan SC, Khairullah AR. (2024). Detection of the chuA gene encoding the invasive enterohemorrhagic species Escherichia coli 0157:H7 using qPCR in horse feces samples on Sumbawa Island, Indonesia. Open Vet J, 14(4), 1051-1058. https://doi.org/10.5455/OVJ.2024.v14.i4.12

Publication

Open veterinary journal
ISSN: 2218-6050
NlmUniqueID: 101653182
Country: Libya
Language: English
Volume: 14
Issue: 4
Pages: 1051-1058

Researcher Affiliations

Kholik, Kholik
  • Department of Veterinary Public Health, Faculty of Veterinary Medicine, Universitas Pendidikan Mandalika, Mataram, Indonesia.
Sukri, Akhmad
  • Departement of Biology Education, Universitas Pendidikan Mandalika, Mataram, Indonesia.
Riwu, Katty Hendriana Priscilia
  • Department of Veterinary Public Health, Faculty of Veterinary Medicine, Universitas Pendidikan Mandalika, Mataram, Indonesia.
Kurniawan, Shendy Canadya
  • Master Program of Animal Sciences, Department of Animal Sciences, Specialisation in Molecule, Cell and Organ Functioning, Wageningen University and Research, Wageningen, The Netherlands.
Khairullah, Aswin Rafif
  • Research Center for Veterinary Science, National Research and Innovation Agency (BRIN), Bogor, Indonesia.

MeSH Terms

  • Animals
  • Escherichia coli Infections / veterinary
  • Escherichia coli Infections / microbiology
  • Escherichia coli O157 / isolation & purification
  • Escherichia coli O157 / genetics
  • Escherichia coli Proteins / genetics
  • Feces / microbiology
  • Horse Diseases / microbiology
  • Horse Diseases / diagnosis
  • Horses
  • Indonesia
  • Real-Time Polymerase Chain Reaction / methods
  • Real-Time Polymerase Chain Reaction / veterinary
  • Bacterial Outer Membrane Proteins / genetics
  • Receptors, Cell Surface / genetics

Conflict of Interest Statement

There is no conflict of interest in this study.

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Citations

This article has been cited 1 times.
  1. Kholik K, Sukri A, Priscilia Riwu KH, Ayu IW, Dewi IN. Molecular detection of the Escherichia coli heme-utilization gene A virulence factor in E. coli isolated from the feces of horses in Sumbawa island, Indonesia.. Vet Res Forum 2025;16(6):325-330.
    doi: 10.30466/vrf.2024.2039823.4421pubmed: 40726538google scholar: lookup
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