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| Product Name | HiSens TaqMan Multiplex qPCR Mix (UDG Plus) |
| Catalog No. | RESK-0133 |
| Description | The HiSens TaqMan Multiplex qPCR Mix (UDG Plus) is a multiplex real-time qPCR premix built on a new-generation antibody-based hot-start Taq DNA polymerase. Leveraging a dedicated enzyme-engineering platform, researchers performed structure-guided, all-round directed evolution of Taq DNA polymerase - combining sequence alignment of enzymes from multiple sources with site-directed mutagenesis, random-mutation directed screening, stress testing and enzyme fragment recombination - to obtain a series of superior mutants. High-throughput multidimensional screening across thermostability, affinity, polymerization activity and exonuclease activity, with screening pressure further optimized for direct-amplification application scenarios, yielded a Taq DNA polymerase with strong inhibitor tolerance and a fast synthesis rate. Built on this engineered enzyme, the qPCR reaction system was optimized through extensive testing with real clinical samples and iterative system adjustment and validation, resulting in a qPCR premix dedicated to African swine fever virus (ASFV) detection. |
| Intended Use | For research and veterinary diagnostic assay development: rapid real-time qPCR detection of African swine fever virus (ASFV) nucleic acid, e.g., in nucleic acids extracted by large-scale magnetic-bead methods from pig-farm samples. |
| Principle / Technology | Antibody-based hot-start Taq DNA polymerase: enzyme activity is blocked at low temperature and released upon heat activation, minimizing nonspecific amplification. The Taq polymerase was obtained by directed evolution (site-directed mutagenesis, random-mutation screening, stress testing and fragment recombination) followed by high-throughput screening for thermostability, affinity, polymerization and exonuclease activities, conferring strong inhibitor tolerance and fast extension speed suited to direct-amplification applications. Carryover contamination control: the dUTP/UDG system degrades dU-containing carryover PCR products, effectively preventing aerosol contamination. |
| Detection Method | TaqMan probe-based real-time quantitative PCR (qPCR); multiplex-capable. |
| Sample Type | ASFV nucleic acid extracted from real clinical samples (e.g., pig-farm samples processed by large-scale magnetic-bead extraction); validated on 45 ASFV clinical specimens. |
| Performance Range / Specifications | Optimized adaptability: system optimized for ASFV nucleic acid detection; well suited to mainstream real-time PCR instruments. High sensitivity: in testing of over one hundred clinical samples, detection rate and sensitivity exceed those of comparable products; superior low-concentration detection and higher fluorescence values. Fast: results in 20 minutes. Stable full premix: performance stable after 7 days at 37 °C with ASFV primers/probes premixed, and after 20 freeze-thaw cycles. dUTP/UDG anti-contamination system effectively prevents aerosol contamination. |
| Sensitivity / LOD | In testing of ASFV nucleic acid serially diluted to limit-of-detection concentrations, the mix showed better sensitivity and earlier Ct values than comparable products; high detection rate and superior low-concentration detection demonstrated across 45 clinical samples. |
| Specificity | No nonspecific amplification observed after heat-accelerated storage of the full premix; antibody hot-start design minimizes nonspecific products. |
| Reaction Conditions / Protocol | Recommended reaction setup: 25 uL total reaction volume with 5 uL template. Cycling program: 95 °C for 10 sec; 40 cycles of 95 °C for 1 sec and 60 °C for 5 sec; total run time approximately 20 minutes. |
| Components / Formulation | Engineered antibody-based hot-start Taq DNA polymerase with strong inhibitor tolerance and fast synthesis rate. Optimized qPCR reaction buffer system. dUTP/UDG carryover contamination control system. |
| Storage Conditions | Store at -25 to -15 °C. |
| Shelf Life | 18 months. |
| Package Specifications | 50 T; 100 T; 1,000 T; 10,000 T |
| Product Form | Liquid premix. |
| Quality Control | Reaction system optimized and validated through extensive testing with over one hundred real clinical samples; full-premix stability confirmed by accelerated and freeze-thaw testing. |
| Expiration Date / Stability | Valid for 18 months when stored at -25 to -15 °C. Full premix containing ASFV primers/probes shows no significant differences in peak shape, Ct value or detection rate after 7 days at 37 °C versus storage at -20 °C, with no nonspecific amplification after heat acceleration; stable through 20 freeze-thaw cycles. |
| Compatibility | Optimized for ASFV detection; compatible with mainstream real-time PCR instruments. |
| Application Notes / Precautions | Recommended 25 uL reaction volume with 5 uL template; nucleic acid prepared by magnetic-bead extraction is recommended. The dUTP/UDG anti-contamination system is built in; include a UDG digestion step as required by the protocol. |
For research use only, not for clinical use.
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