Related Experiment Video
Updated: Jun 12, 2026
Performing and Processing FNA of Anterior Fat Pad for Amyloid
Published on: October 30, 2010
Real-time PCR detection of Enterococcus faecalis associated with amyloid arthropathy
A Petersen1, M Bisgaard, H Christensen
1Department of Veterinary Disease Biology, Faculty of Life Sciences, University of Copenhagen, Copenhagen, Denmark.
Aims:
To develop a RT-PCR method for detection of the multilocus sequence type 82 of Enterococcus faecalis associated with amyloid arthropathy (AA) in layers.
Methods And Results:
Bacteria were selected from lesions including AA in layers. The primers were designed based on the phosphate ATP binding cassette transporter (pstS) and xanthine phosphoribosyltransferase (xpt) genes and first tested against three isolates with known base pairs at the specific sites. Subsequently, 12 isolates were selected from our collection by one researcher, and RT-PCR was performed blinded. The sequence type (ST) was then confirmed by multilocus sequence analysis. Two single-nucleotide polymorphisms in the pstS and xpt genes allowed an unambiguous identification of ST82. As an alternative to DNA extraction, a boiling method for release of DNA from cells was used.
Conclusions:
The real-time PCR targeting ST82 enables rapid screening of Ent. faecalis cultured from suspect cases with results available after a few hours, much faster than multilocus sequence typing and pulse field gel electrophoresis.
Significance And Impact Of The Study:
The new method allows a rapid screening of isolates with results available after only few hours. This RT-PCR method could be a useful tool for molecular epidemiological studies on the spread of arthropathic and amyloidogenic Ent. faecalis within and between birds more efficiently.
Insights
A new real-time PCR method rapidly detects Enterococcus faecalis ST82, a cause of amyloid arthropathy in poultry. This faster screening tool aids in tracking the spread of this specific bacterial strain.
Area of Science:
- Veterinary Microbiology
- Molecular Diagnostics
- Poultry Pathology
Background:
- Enterococcus faecalis is associated with amyloid arthropathy (AA) in layers.
- Multilocus sequence typing (MLST) is used to characterize bacterial strains.
- Efficient detection methods are needed for epidemiological studies.
Purpose of the Study:
- To develop a rapid real-time PCR (RT-PCR) method for detecting Enterococcus faecalis multilocus sequence type 82 (ST82).
- To identify a specific bacterial strain linked to amyloid arthropathy in poultry.
Main Methods:
- Primers were designed targeting the phosphate ATP binding cassette transporter (pstS) and xanthine phosphoribosyltransferase (xpt) genes.
- RT-PCR was performed on 12 Enterococcus faecalis isolates.
- A boiling method for DNA extraction was evaluated as an alternative to standard DNA extraction.
Main Results:
- Two single-nucleotide polymorphisms in the pstS and xpt genes enabled unambiguous identification of ST82.
- The RT-PCR method provided results within hours.
- The boiling method for DNA release was effective.
Conclusions:
- The developed RT-PCR method allows for rapid screening of Enterococcus faecalis ST82.
- This technique is significantly faster than traditional methods like MLST and pulse field gel electrophoresis.
- The RT-PCR assay is a valuable tool for molecular epidemiological studies on arthropathic and amyloidogenic Enterococcus faecalis in poultry.
More Related Videos
07:27Methods for Detecting Cytotoxic Amyloids Following Infection of Pulmonary Endothelial Cells by Pseudomonas aeruginosa
Published on: July 12, 2018
10:04Imaging Amyloid Tissues Stained with Luminescent Conjugated Oligothiophenes by Hyperspectral Confocal Microscopy and Fluorescence Lifetime Imaging
Published on: October 20, 2017
