Related Experiment Video
Updated: Aug 29, 2025
06:34
Identification of Rare Bacterial Pathogens by 16S rRNA Gene Sequencing and MALDI-TOF MS
Published on: July 11, 2016
18.4K
A New Specific DNA Target Sequence for Identification of Staphylococcus epidermidis using Modified Comparative
Reza Khoshbakht1,2, Hosna Zare1,2, Reza Kamali Kakhki1,2
1Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.
Avicenna Journal of Medical Biotechnology
|September 5, 2022
Summary
A new, highly specific DNA target sequence (Se400) was identified for detecting Staphylococcus epidermidis, a common cause of prosthesis infections. This finding is crucial for developing more accurate diagnostic tests like Nucleic Acid Amplification Tests (NAAT).
Area of Science:
- Microbiology
- Genomics
- Molecular Diagnostics
Background:
- Staphylococcus epidermidis is a primary cause of infections associated with medical prostheses.
- Current diagnostic methods, including Nucleic Acid Amplification Tests (NAAT), require more specific targets for accurate detection.
- The lack of complete specificity in existing DNA targets necessitates the identification of novel, highly specific sequences.
Purpose of the Study:
- To identify a novel, 100% specific DNA target sequence for Staphylococcus epidermidis.
- To develop and evaluate a Polymerase Chain Reaction (PCR) method based on the identified target sequence.
- To improve the diagnostic accuracy of Staphylococcus epidermidis detection, particularly in the context of prosthesis infections.
Main Methods:
- Modified comparative genomic analysis was employed to discover the most specific DNA target.
- A PCR assay was designed and optimized for the selected target sequence.
- Analytical specificity and detection limits were determined using genomic DNA from a panel of Staphylococcus epidermidis strains and other bacteria.
Main Results:
- A 400 bp sequence (Se400) from Staphylococcus epidermidis ATCC 14990 demonstrated 100% sequence similarity with Staphylococcus epidermidis genomes and no similarity with other bacterial genomes.
- The Se400-based PCR assay exhibited a detection limit of 10 fg, equivalent to approximately 4 copies of Staphylococcus epidermidis genomic DNA per microliter.
- The PCR assay successfully generated a 264 bp amplicon for all tested Staphylococcus epidermidis samples while yielding negative results for non-Staphylococcus epidermidis bacteria.
Conclusions:
- The Se400 sequence represents a highly specific and reliable target for the molecular detection of Staphylococcus epidermidis.
- This finding has significant implications for the development of advanced diagnostic tools for Staphylococcus epidermidis infections.
- The validated Se400 target can enhance the specificity and sensitivity of NAAT-based diagnostic platforms.
Related Concept Videos
01:29Modern Molecular Taxonomy
109
Advancements in molecular biology have revolutionized the identification and characterization of bacteria, with multiple methods leveraging DNA sequencing for enhanced precision. As sequencing technologies improve and costs decline, these approaches are increasingly used in clinical, environmental, and evolutionary studies.Multilocus Sequence Typing (MLST) examines several housekeeping genes, essential chromosomal genes encoding cellular functions, to distinguish strains. Approximately...
109
01:20Applications of Molecular Taxonomy
76
Molecular taxonomy has revolutionized the understanding and classification of bacteria, providing precise insights into their diversity, evolutionary relationships, and ecological roles. By utilizing molecular techniques such as DNA sequencing and fingerprinting, researchers have made significant strides in various fields related to bacterial studies.Resolving Taxonomic AmbiguitiesMolecular taxonomy has been instrumental in distinguishing closely related bacterial species initially thought to...
76
01:57Sanger Sequencing
756.3K
DNA sequencing is a fundamental technique that is routinely used in the biological sciences. This method can be applied to a range of questions at different scales - from the sequencing of a cloned DNA fragment or the study of a mutation in a gene up to whole-genome sequencing. However, despite the widespread use of sequencing today, it was not until 1977 that Fredrick Sanger and his collaborators developed the chain-termination method to decode DNA sequences. It relies on the separation of a...
756.3K
01:28Methods of Classification and Identification
134
Bacterial identification relies on a diverse array of techniques to classify and understand microorganisms, each tailored to uncover specific characteristics. Traditional morphological approaches, while still valuable, are limited for closely related or structurally simple organisms. Modern methods integrate biochemical, serological, genetic, and advanced molecular tools to achieve greater accuracy.Morphological and Biochemical TechniquesMorphological characteristics, such as cell shape and...
134