Keywords:
Low residue,Pre-mixed,Universal Multi-Target, One-Tube, qPCR, High Sensitivity.
Abstract:
The new third-generation ultra-low residue qPCR mix, pre-mixed to ensure high sensitivity, supports a rapid 30-minute protocol and excels in multiplex amplification across diverse scenarios.
Background：
Commercial molecular enzymes, primarily produced via recombinant expression in engineered strains (e.g., E. coli), often contain residual host genomic DNA. Factors such as the preparation environment and human contamination further increase the likelihood of residual DNA in these enzymes. This may lead to inaccurate quantification of host nucleic acid residuals in quality control products, posing safety risks to biopharmaceutical manufacturing. During pathogen detection, contaminated background bacterial nucleic acids could overwhelm low-abundance target nucleic acids or be co-detected with them, compromising test accuracy.
Total Solution
To tackle interference from background bacteria and host nucleic acid residuals in pathogen detection, YEASEN Biotech has developed a unique ultra-low residue control technology. We established UCF.ME, an ultra-clean molecular enzyme production base adhering to GMP standards, with strict compliance to the ISO 13485 quality management system across all stages. This includes rigorous control over personnel, machinery, materials (reagents and consumables), processes, environment, and standards. By integrating effective technical measures, a clean production environment, and stringent quality control, we enable large-scale production of various UCF.METMultra-low residual molecular enzymes.
Hieff UniconTM Pure Pro U+ qPCR Mix（One Tube）（Cat#16713ES）is a meticulously crafted universal qPCR mix by YEASEN Biotech, which features low host DNA residue. This product is based on a new generation of universal pre-mixed qPCR mix (Cat#16713ES) that effectively controls the residual genomic DNA of the host (E. coli), while also controlling the residual levels of common pathogenic bacteria such as Pseudomonas aeruginosa and human-derived contaminants. Since this pre-mixed solution (Cat#16713ES) not only effectively controls the level of pathogen residue but also addresses the issues of stability in multiple detection and target versatility, it enhances key performance indicators such as the sensitivity and specificity of the reagent. It significantly improves detection efficiency and can be widely used in application scenarios such as pathogenic microorganism detection.
Product Advantages
- Low background residue – No detection of Escherichia coli, Pseudomonas aeruginosa and human-derived genomic DNA residue in 48 wells.
- Exceptional pre-mixed stability: Stable at 37°C for up to 14 days, at 4°C for up to 28 days, and after 50 freeze-thaw cycles.
- Excellent reagent versatility – Compatible with a variety of multiplex primer-probe sets (validated for over 40 different types of targets), with excellent amplification performance.
- High sensitivity and specificity – The sensitivity is capable of detecting 250 copies/mL, with specificity ensuring no peaks in 48 negative wells.
- Supports rapid protocols – Compatible with rapid protocols, results can be obtained in just 30 minutes.
- dUTP/UDG anti-contamination system – Incorporates a dUTP/UDG anti-contamination system to effectively prevent aerosol contamination.
- Compatibility with multiple platforms and systems – Compatible with platforms such as Bio-Rad CFX96, ABI Q5, 7500, Slan, Tianlong, and others.
Performance Highlights
1、Ultra-low background bacterial residue
Based on the assessment of reagent background bacterial residue levels, the use of YEASEN’s ultra-low residue universal pre-mixed qPCR mix (Cat#16713ES) and conventional reagents (without residue control) for NTC(No Template Control) amplification in an Escherichia coli (E. coli) system, with 16713ES also undergoing NTC amplification in both Pseudomonas aeruginosa and human-derived genomic DNA systems. The results show that Cat#16713ES exhibits no amplification in 48 wells of NTC, indicating negligible background DNA residue (E. coli, P. aeruginosa, and human DNA) in the reagent.
2、Outstanding stability of the fully pre-mixed pre-mix
Based on the investigation of the stability of the pre-mixed reagent, Cat#16713ES was treated in three ways (4°C, 37°C, and freeze-thaw). The experimental groups were treated at 4°C and 37°C for 7 days, respectively, and subjected to 50 freeze-thaw cycles. The stability was evaluated against the control group (stored at -20°C) under four multiplex amplification systems (Groups 1, 2, and 4 are 4-plex amplification systems, and Group 3 is a 3-plex amplification system), with some amplification groups additionally assessed for stability after 14 days of acceleration at 37°C. The results demonstrated that the Ct value deviation (ΔCt value) between the experimental and control reagents was within ±0.5, and the fluorescence value deviation was within 15%, indicating that YEASEN’s pre-mixed reagent has excellent stability in multiplex amplification.
3、Excellent reagent versatility
Based on the investigation of the reagent’s versatile application scenarios, YEASEN (Cat#16713ES) was used alongside several conventional reagents to validate amplification performance under different target scenarios. The results demonstrated that the product exhibits good amplification performance across a variety of targets including pathogens, human sources, and plants, indicating a high level of reagent versatility.
4、Outstanding Amplification Sensitivity
Based on an assessment of reagent sensitivity, an evaluation was conducted on the detection rate of Cat#16713ES in a multiplex amplification system (4-plex) using samples at ultra-low concentrations (250 copies/mL). The results indicate that YEASEN's Cat#16713ES exhibits excellent amplification sensitivity, with a limit of detection of at least 250 copies/mL.
5、Compatibility with multiple rapid protocols
Based on the investigation of the amplification performance of the reagent’s rapid protocol, seven gradient samples (1~10^6 copies/μL) were validated using both conventional (1 hour and 6 minutes) and rapid protocols (30 minutes) on the same quantitative PCR instrument (ABI Q5). The results demonstrated that the amplification efficiency of the rapid protocol remained above 99%, and the Ct value deviation (ΔCt value) between the conventional and rapid protocols was within ±0.5, with a fluorescence value deviation within ±10%, and the amplification curves displayed an “S” shape.
IVD Detection Reagent Sharing
As a supplier of life science tools, YEASEN Biotech has been providing high-quality and cost-effective raw materials to IVD companies for a long time. In addition to universal fully pre-mixed qPCR/RT-qPCR master mixes, we offer a variety of other qPCR/RT-qPCR detection reagents, helping IVD enterprise customers jointly address the efficient detection of human pathogens and animal diseases.
|
Product Type |
Product Function |
Product Name |
Item Number |
|
qPCR mix |
Brand-new 3rd generation universal fully pre-mixed, high-sensitivity supports rapid testing |
16710ES |
|
|
High-sensitivity universal (5× concentration) |
Hieff UniconTM Universal TaqMan Multiplex qPCR Master Mix（UDG plus） |
13891ES |
|
|
2nd generation rapid fully pre-mixed |
Hieff UniconTM Superpro Ⅰ TaqMan qPCR Master Mix（UDG plus） |
11827ES |
|
|
Host-free ultra-low residual |
Hieff UniconTM Purepro Ⅰ TaqMan qPCR Master Mix（UDG plus） |
11853ES |
|
|
High-sensitivity universal lyophilizable |
Hieff UniconTM Universal TaqMan Multiplex qPCR Master Mix（UDG plus） |
11893ES |
|
|
RT-qPCR mix |
High-sensitivity universal rapid |
HifairTM C203P1 Multiplex One Step RT-qPCR Probe Kit（UDG Plus） |
16630ES |
|
fully pre-mixed universal |
HifairTM V Multiplex One Step RT-qPCR Probe Kit（UDG Plus） |
11899ES |
|
|
Universal rapid lyophilizable |
11831ES |