High-Quality Universal RT-qPCR Reagents — Empowering Molecular Diagnostic Kit Development

Breakthrough Technologies: Intelligent Boost in Efficiency and Stability

Through precise engineering of core reaction conditions and intelligent formulation design, we have significantly elevated the performance of our RT-qPCR kits. From dynamic buffer optimization to enzyme purity control and surfactant synergy, each component is fine-tuned to work in harmony—delivering faster amplification, higher sensitivity, and unmatched stability across storage and transport conditions. This intelligent process integration empowers next-generation diagnostic development with confidence and consistency.

Process Control and Optimization: Empowered by Smart Manufacturing

Yeasen Biotech, with years of experience in the field of enzyme raw materials, has been providing high-quality and cost-effective molecular diagnostic PCR raw materials to IVD companies, third-party testing institutions, and research organizations.

Critical Process Parameter (CPP) Monitoring

• Core Parameters: Mixing uniformity (CV < 3%), pH control (8.8 ± 0.1), Ionic strength (e.g., Na⁺ 50 mM ± 5%)

Intelligent Quality Control System

• AI-Powered Monitoring: Predicts deviations (e.g., temperature, humidity) and adjusts parameters in real time.
• NIR Spectroscopy: Real-time dNTP concentration monitoring with ±0.5 μM accuracy.
• Case Study: Implementation of AI-based QC at Company M*I reduced defect rate from 2% to 0.5%.

Continuous Process Verification & Optimization

• Batch Monitoring: 30 batches tested quarterly with inter-batch Ct value variation < 0.5.
• Closed-Loop Feedback: Real-time user feedback refines parameters (e.g., reagent composition, primer concentrations).
• Case Study: After 5 process iterations, Daan Gene’s HPV kit saw clinical concordance increase from 95% to 99.5%.

 

Universal High-Performance RT-qPCR Master Mix, Fast program support

HifairTM V2 Multiplex One Step RT-qPCR Probe Kit（UDG Plus） Cat#16630ES

• Strong versatility: suitable for one-step RT-qPCR detection of nucleic acids from multiple species such as pathogenic microorganisms, human genomes, and plants;
• High sensitivity: The carefully optimized buffer system improves the detection sensitivity of low-concentration templates up to 250 copies/mL;
• dUTP/UDG anti-pollution system: dUTP/ UDG anti-pollution system is introduced to degrade aerosol pollution of products efficiently, reduce false positive, and ensure authentic results;
• Support fast program: compatible with fast program, 30min can produce results.
• Super stability: Stable performance after 14 days at 37°C, long-distance transport, and 10 freeze-thaw cycles.

Partial performance display

1. Excellent amplification performance

Figure 1. Amplification efficiency test

16630 linear range results: amplification efficiency meets 90%-110%, and the template range when the slope of the standard curve is 3.3 is 106 copies/μL to 10-2 copies/μL.

There are 8 dilution gradients in total, and the lowest concentration of Supplier A* was not detected.

2.High sensitivity

Figure 2. Sensitivity comparison test 

Using the parainfluenza quadruple primer probe, 20 replicate wells of parainfluenza T7-RNA plasmid were tested, and the results showed that the sensitivity of pathogen quadruple target amplification could reach 0.25 copies/μL.

3. High specificity

Figure 3. Specificity test 

Evaluation Scheme:
Human blank samples (e.g., negative oral swabs) were used to test specificity of pathogen primers, and ultrapure water was used to test specificity of human primers. The assay included a 4-plex pathogen panel and a 4-plex human target panel.

Results:
No nonspecific amplification peaks were observed in 24 replicates of pathogen targets with negative oral swab lysate, nor in 24 replicates of human targets with ultrapure water.

4. Support for fast programs

Figure 4. Comparison of Different Brands under Rapid PCR Program

Under the Q5 fast program, reagent 16630 demonstrated superior detection performance for RSVA, RSVB, and HRVA, HRVB, HRVC compared to reagents 11214 and 13650. The 16630ES reagent is fully compatible with these fast protocols.

5. High Inhibitor Tolerance

Figure 5. Inhibitor tolerance testing

16630 and supplier-A fully tolerated 4% ethanol, and 4% whole blood inhibited both to a similar extent.

6. Stability Test

Figure 6. Thermal accelerated stability testing(A) and Freeze-thaw stability(B)

(A) Accelerated Stability Test:
Enzyme and buffer components of the single-component 16630 reagent (two-tube format: buffer and enzyme) were stored at 4°C and 37°C for 14 days. Under quadruplex pathogen and human target detection, amplification performance showed no significant difference compared to −20°C storage, indicating qualified stability.

(B) Freeze–Thaw Stability Evaluation:
Amplification performance was evaluated after 10 freeze–thaw cycles using dry ice and −20°C storage conditions. Both pathogen and human quadruplex targets were tested. Results showed qualified stability under both freeze–thaw conditions for all target sets.

 

Product Overview