In the realm of molecular biology, the quest for efficient and precise nucleic acid amplification techniques has been a driving force behind numerous groundbreaking innovations. Among these, Recombinase Polymerase Amplification (RPA) and Polymerase Chain Reaction (PCR) stand out as two formidable methodologies, each with its unique strengths and applications.
Principle:
- PCR: PCR involves repeated cycles of denaturation, annealing, and extension using a thermostable DNA polymerase enzyme.
- RPA: RPA is an isothermal amplification technique that uses recombinase proteins to facilitate primer binding and strand displacement, followed by polymerase-mediated DNA synthesis.
Temperature Requirements:
- PCR: Requires cycling through different temperature stages (denaturation, annealing, extension) typically ranging from 50°C to 95°C.
- RPA: Operates at a constant, lower temperature (usually around 37-42°C), eliminating the need for thermal cycling equipment.
Speed:
- PCR: Generally takes around 1-2 hours due to cycling through multiple temperature stages.
- RPA: Typically faster than PCR, with reaction times ranging from 10 to 60 minutes, making it advantageous for rapid diagnostics.
Sensitivity:
- PCR: Highly sensitive and capable of detecting low concentrations of DNA.
- RPA: Also highly sensitive and capable of detecting low copy numbers of target DNA, comparable to PCR.
Equipment:
- PCR: Requires a thermal cycler to control temperature cycles.
- RPA: Does not require a thermal cycler, making it more accessible, especially for point-of-care testing or field applications.
Applications:
- PCR: Widely used in research, clinical diagnostics, forensics, and molecular biology applications.
- RPA: Also applicable in diagnostics, particularly in resource-limited settings or fieldwork due to its rapid and equipment-independent nature. It's also gaining popularity in environmental monitoring and food safety testing.
Ease of Use:
- PCR: Requires precise temperature control and cycling conditions, often necessitating trained personnel and specialized equipment.
- RPA: Simplified protocol and isothermal conditions make it more user-friendly, potentially enabling use by non-specialists in various settings.
As a summary, RPA, a relatively recent addition to the molecular biologist's toolkit, has rapidly gained prominence due to its isothermal nature and rapid amplification capabilities. Unlike PCR, which relies on thermal cycling to denature, anneal, and extend DNA strands, RPA operates at a constant temperature, typically around 37-42°C. This feature eliminates the need for sophisticated thermal cycling equipment, making RPA particularly attractive for point-of-care testing (POCT) and field applications where resources may be limited.
Furthermore, RPA's reliance on recombinase proteins to facilitate primer binding and strand displacement, followed by polymerase-mediated DNA synthesis, enables swift amplification of target sequences. The simplicity of the protocol and the speed of the reaction, typically completing within 10 to 20 minutes, make RPA an invaluable tool in scenarios requiring rapid diagnostics and screening.
Despite these advantages, PCR remains the gold standard in many molecular biology laboratories due to its well-established protocols, high sensitivity, and versatility. However, RPA's emergence has undoubtedly expanded the landscape of nucleic acid amplification techniques, offering novel solutions to old challenges.
Introducing the RPA Multiplex Fluorescent Lyophilized Kit (Gen-4)
In line with the relentless pursuit of innovation, SBS Genetech proudly presents the RPA Multiplex Fluorescent Lyophilized Kit (Gen-4), heralding a groundbreaking chapter in nucleic acid research and POCT, combining the advantages of both RPA and PCR. This innovative kit represents the convergence of cutting-edge multiplex RPA technology with the convenience of lyophilized reagents, setting new standards for efficiency, sensitivity, and ease of use.
With our exclusive domestically developed multiplex RPA reagents, the Gen-4 kit empowers simultaneous amplification of 2-4 targets within a mere 10 to 20 minutes. This unparalleled capability supports the direct amplification of DNA/RNA with exceptional specificity and sensitivity, achieving a remarkable minimum detection limit of 400 copies/mL.
Moreover, the Gen-4 kit features the innovative dUTP-UDG anti-aerosol contamination system, engineered to eradicate aerosol contamination of 10 copies or more. We offer a choice between two contamination prevention systems, tailored to meet the needs of fluorescence and CRISPR technologies alike.
Leveraging advanced freeze-drying technology, our product is presented in lyophilized form, ensuring significantly enhanced thermostability. This allows for shipment at room temperature, thereby substantially reducing shipping costs while maintaining product integrity.
Features of the RPA Multiplex Fluorescent Lyophilized Kit (Gen-4)
- Multiplex Recombinase Polymerase Amplification (RPA): Exclusive domestically developed multiplex RPA reagents enable simultaneous amplification of 2-4 targets, with testing completed within 10-20 minutes, supporting direct amplification of DNA/RNA.
- High Specificity and Sensitivity: Utilizing the latest formulation processes to effectively reduce nonspecific amplification; achieving a lowest detection limit of 400 copies/mL.
- Aerosol Contamination Prevention: Incorporating a novel dUTP-UDG aerosol contamination prevention system; capable of eliminating 10 copies of aerosol contamination. The contamination prevention system can be tailored as required:
- Product degradation upon amplification completion (suitable for fluorescence-based methods)
- Product degradation within aerosols (CRISPR).
- Room Temperature Shipment: With advanced freeze-drying technology, RPA Multiplex Fluorescent Lyophilized Kit (Gen-4) is provided in lyophilized form with much higher thermostability, which can be shipped at room temperature, greatly saving the shipping cost.
Based on our leading RPA technology, we have developed a series of reagent kits to enrich the field of POCT.
- Tuberculosis Complex Rapid Amplification Kit (with internal reference)
- Type A Influenza Virus Rapid Amplification Kit (with internal reference)
- Type B Influenza Virus Rapid Amplification Kit (with internal reference)
- Type A/B Influenza Virus Rapid Amplification Kit (with internal reference)
- Mycoplasma pneumoniae Rapid Amplification Kit (with internal reference)
- Cytomegalovirus A/B Rapid Amplification Kit (with internal reference)
- Chlamydia trachomatis/Neisseria gonorrhoeae Combined Rapid Amplification Kit (with internal reference)
- Human Immunodeficiency Virus Rapid Amplification Kit (with internal reference)
- Chlamydia trachomatis Rapid Amplification Kit (with internal reference)
- Neisseria gonorrhoeae Rapid Amplification Kit (with internal reference)
- Human Papillomavirus 15 High-Risk Type Rapid Amplification Kit (with internal reference)
- African Swine Fever Virus Rapid Amplification Kit (with internal reference)
- Rabies Virus Rapid Amplification Kit (with internal reference)
- Brucella Rapid Amplification Kit (with internal reference)
- Canine Distemper Virus Rapid Amplification Kit (with internal reference)
- Canine Parvovirus Rapid Amplification Kit (with internal reference)
- Canine Adenovirus Rapid Amplification Kit (with internal reference)
Each kit offers real-time fluorescence or paper strip-based detection, with a lowest detection limit of 500 copies/mL and includes an internal reference for accurate results. These kits cater to various samples and target pathogens, further enhancing the capabilities of POCT.
With SBS Genetech's leading RPA technology, we aim to provide innovative solutions for rapid and reliable molecular diagnostics in diverse settings.