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Related Concept Videos

Labeling DNA Probes03:31

Labeling 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...
Southern Blot02:57

Southern Blot

Agarose gel electrophoresis is very useful in separating DNA fragments by size. Running a DNA ladder containing fragments of the known length alongside the sample helps determine the approximate length of the sample DNA fragments. However, additional steps are needed to verify the sequence identity of the sample DNA fragments.
Denatured DNA fragments must be transferred onto a carrier membrane from the gel to make it accessible to a probe - a small ssDNA fragment complementary to the target DNA...
FISH - Fluorescent In-situ Hybridization02:07

FISH - Fluorescent In-situ Hybridization

Fluorescence in situ hybridization, or FISH, was developed in the early 1980s and has quickly become one of the most widely used techniques in cytogenetics. Labeled probes are used to bind complementary DNA or RNA sequences on a chromosome or in a region within a cell. Earlier, the probes could only be obtained by cloning or reverse transcription of a DNA template. Currently, the probe oligonucleotides can be synthesized synthetically. Additionally, with the advancement of optical techniques,...

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Related Experiment Video

Updated: Jul 13, 2026

Split Hybridization Probe Utilizing a DNA Fluorescent Light-up Aptamer as a Signal Reporter for Sequence-Specific Nucleic Acid Analysis
07:10

Split Hybridization Probe Utilizing a DNA Fluorescent Light-up Aptamer as a Signal Reporter for Sequence-Specific Nucleic Acid Analysis

Published on: July 8, 2025

LAMP-Based Two-DNA-Fragment Fusion and Its Application in Nucleic Acid Detection.

Qinghong Zhou1, Bingyan Xu1, Yi Wang1

  • 1Key Laboratory of Laboratory Medicine, Ministry of Education, School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, Wenzhou 325035, China.

ACS Synthetic Biology
|July 11, 2026
PubMed
Summary

A new fusion LAMP method enables DNA fragment assembly and, with CRISPR/Cas13a, creates a Fusion LAMP-Coupled CRISPR/Cas13a (FLCC) platform for accurate pathogen detection, like methicillin-resistant Staphylococcus aureus (MRSA). This AND-gate system only signals when two targets are present.

Keywords:
CRISPR/Cas13aDNA fragment fusionMRSAfusion LAMPnucleic acid detection

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Published on: November 15, 2017

Area of Science:

  • Molecular Biology
  • Biotechnology
  • Diagnostic Assays

Background:

  • Loop-mediated isothermal amplification (LAMP) generates DNA intermediates suitable for fragment assembly.
  • CRISPR-Cas13a systems offer precise nucleic acid detection capabilities.

Purpose of the Study:

  • To develop a novel isothermal DNA-fusion technique (fusion LAMP) for assembling two DNA fragments.
  • To establish an "AND-gate" nucleic acid detection platform (FLCC) by integrating fusion LAMP with CRISPR/Cas13a for concurrent dual-target detection.
  • To validate the FLCC platform's efficacy in detecting methicillin-resistant Staphylococcus aureus (MRSA).

Main Methods:

  • Developed fusion LAMP, an isothermal strategy for fusing two independent DNA fragments in one reaction.
  • Integrated fusion LAMP with CRISPR/Cas13a to create the FLCC platform, triggering fluorescence upon fusion product formation.
  • Applied FLCC to detect methicillin-resistant Staphylococcus aureus (MRSA) genomic DNA.

Main Results:

  • Achieved a limit of detection of 10 copies/μL for MRSA genomic DNA using the FLCC platform.
  • Demonstrated high specificity with no cross-reactivity against closely related bacterial strains.
  • Successfully validated the FLCC platform's performance using 19 clinical isolates of MRSA.

Conclusions:

  • The FLCC platform provides a sensitive and specific method for identifying pathogens by detecting the simultaneous presence of two targets.
  • Fusion LAMP coupled with CRISPR/Cas13a offers a promising diagnostic approach for complex genetic target detection and pathogen identification.
  • The developed platform presents a simple, accurate, and novel diagnostic tool for clinical applications, particularly for MRSA detection.