Related Experiment Video
Updated: Jul 2, 2026
07:59
Rapid and Specific Detection of Acinetobacter baumannii Infections Using a Recombinase Polymerase Amplification/Cas12a-based System
Published on: April 25, 2025
Rapid detection and identification of a pathogen's DNA using Phi29 DNA polymerase
Yun Xu1, Simon Gao, John F Bruno
1Department of Medicine, T-16 Room 027, State University of New York at Stony Brook, Stony Brook, NY 11794-8154, USA. jbruno@notes.cc.sunysb.edu
Biochemical and Biophysical Research Communications
|August 30, 2008
Summary
This study introduces a new method for rapidly detecting and identifying circular DNA from infectious agents like Borrelia burgdorferi, even in complex samples. This cultivation-free technique offers a sensitive approach for biothreat agent detection.
Area of Science:
- Microbiology
- Molecular Biology
- Infectious Diseases
Background:
- Zoonotic pathogens, particularly those spread by insect vectors, pose significant threats to human health.
- Some of these agents are recognized as potential biothreat agents due to their weaponization potential.
Purpose of the Study:
- To develop a novel, cultivation-free method for the rapid, differential detection and identification of circular plasmids in infectious agents.
- To profile genomic DNAs using a new in vitro amplification technique.
Main Methods:
- Utilized multiply-primed rolling circle in vitro amplification for DNA profiling.
- Employed Phi29 DNA polymerase and a two-step priming reaction.
- Characterized circular DNA via DNA sequencing.
Main Results:
- Successfully detected and characterized circular DNA from Borrelia burgdorferi B31.
- Achieved reproducible detection with as little as 25 pg of Borrelia DNA in samples with excess human DNA.
- Demonstrated a cultivation-free method for pathogen DNA detection.
Conclusions:
- The developed method allows for rapid, cultivation-free detection and identification of circular plasmids in infectious agents.
- This technology can be adapted for sensitive detection of specific pathogen DNA in diverse and complex environments.
- Offers a promising tool for identifying known and unknown biothreat agents.
Related Concept Videos
01:24DNA Isolation
DNA isolation protocols can be fast and straightforward or complex and time-consuming depending on the type and quality of DNA required for further processing. For example, plasmid DNA extraction is a bit more complicated than genomic DNA extraction because of the need for an appropriate lysis method to separate plasmid DNA from gDNA during isolation. However, for specific applications, such as long-range DNA sequencing that require a good yield of high- quality DNA samples, we need to follow...
01:25Rapid Identification of Pathogens
MALDI-TOF MS has transformed clinical microbiology by offering a rapid and reliable method for pathogen identification. The traditional approach to microbial identification typically involves time-consuming culture techniques and biochemical tests, which can delay the initiation of appropriate antimicrobial therapy. MALDI-TOF MS avoids these delays by using characteristic ribosomal protein mass patterns of microbial cells, enabling accurate species-level identification within minutes.Principle...
03:31Labeling DNA Probes
DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
01:57Sanger Sequencing
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...
01:32PCR
Overview