DNA detection using recombination proteins, specifically recombinase polymerase amplification (RPA), offers a novel method for amplifying and detecting DNA without the need for complex equipment or thermal cycling. RPA uses isothermal amplification driven by recombinase proteins to target and amplify specific DNA sequences, operating at a constant low temperature. This method is sensitive, specific, and rapid, allowing for the detection of as few as ten copies of genomic DNA. The technology also incorporates a probe-based detection system that enables the identification of DNA targets in a simple sandwich assay, making it suitable for instrument-free DNA testing. RPA works by binding opposing oligonucleotide primers to template DNA and extending them with a DNA polymerase. Recombinase-primer complexes scan double-stranded DNA, facilitating strand exchange at cognate sites. The resulting structures are stabilized by single-stranded DNA binding proteins, preventing primer ejection. Recombinase disassembly allows the 3'-end of the oligonucleotide to be accessible to a strand-displacing DNA polymerase, leading to exponential amplification. The dynamic reaction environment balances the formation and disassembly of recombinase-primer filaments, essential for RPA's efficiency. The technology was tested using a pathogen, methicillin-resistant Staphylococcus aureus (MRSA), demonstrating its ability to detect low-template DNA. RPA was also used to detect multiple genetic markers from complex human genomic DNA, showing its versatility and specificity. The method was further validated using a multiplex approach to detect different MRSA alleles and an internal control in the same reaction. RPA's combination of amplification and detection capabilities makes it a significant advancement in the development of portable and widely accessible nucleic acid-based tests. It can be integrated with lateral-flow dip-stick technology for instrument-free detection, enabling point-of-care testing. The method's simplicity and sensitivity make it suitable for various research and clinical applications, particularly in non-laboratory settings. The study highlights the potential of RPA as a breakthrough in DNA detection technology, offering a cost-effective and accessible solution for nucleic acid testing.DNA detection using recombination proteins, specifically recombinase polymerase amplification (RPA), offers a novel method for amplifying and detecting DNA without the need for complex equipment or thermal cycling. RPA uses isothermal amplification driven by recombinase proteins to target and amplify specific DNA sequences, operating at a constant low temperature. This method is sensitive, specific, and rapid, allowing for the detection of as few as ten copies of genomic DNA. The technology also incorporates a probe-based detection system that enables the identification of DNA targets in a simple sandwich assay, making it suitable for instrument-free DNA testing. RPA works by binding opposing oligonucleotide primers to template DNA and extending them with a DNA polymerase. Recombinase-primer complexes scan double-stranded DNA, facilitating strand exchange at cognate sites. The resulting structures are stabilized by single-stranded DNA binding proteins, preventing primer ejection. Recombinase disassembly allows the 3'-end of the oligonucleotide to be accessible to a strand-displacing DNA polymerase, leading to exponential amplification. The dynamic reaction environment balances the formation and disassembly of recombinase-primer filaments, essential for RPA's efficiency. The technology was tested using a pathogen, methicillin-resistant Staphylococcus aureus (MRSA), demonstrating its ability to detect low-template DNA. RPA was also used to detect multiple genetic markers from complex human genomic DNA, showing its versatility and specificity. The method was further validated using a multiplex approach to detect different MRSA alleles and an internal control in the same reaction. RPA's combination of amplification and detection capabilities makes it a significant advancement in the development of portable and widely accessible nucleic acid-based tests. It can be integrated with lateral-flow dip-stick technology for instrument-free detection, enabling point-of-care testing. The method's simplicity and sensitivity make it suitable for various research and clinical applications, particularly in non-laboratory settings. The study highlights the potential of RPA as a breakthrough in DNA detection technology, offering a cost-effective and accessible solution for nucleic acid testing.