As one of the mainstream solutions for molecular POCT on-site testing, LAMP technology has been widely accepted in various application fields due to its fast reaction speed, high sensitivity, and compatibility with multiple technology routes such as colorimetric, fluorescent, probe, and test strip. However, as our partners continue to expand product applications to areas such as animal testing, pet testing, agricultural and forestry infectious disease and transgenic detection, reproductive health, and respiratory health testing, there is a higher demand for product performance that is more convenient, accurate, and sensitive, which imposes higher requirements on the core material BST
By comprehensively analyzing and summarizing the needs of domestic and foreign customers, we have leveraged our company’s advantages in research and development resources. Starting from several dimensions including molecular construction, adapter research, application analysis and testing, freeze-drying process, and large-scale production amplification, we have completely upgraded the BST enzyme.
What are the characteristics of the new version of BST?
Based on user demands, the new BST V2 features the following characteristics, making it highly resistant and adaptable to the requirements of molecular POCT:
- Fast amplification speed: In the application system, it achieves lower CT values and faster positive results.
- Good sample compatibility: Compatible with various DNA and RNA target detection, low detection limit, facilitating rapid matching of primers for customers.
- Strong inhibition resistance: It tolerates various inhibitory components, making it more suitable for the development of extraction-free amplification systems and expanding the coverage range of customer products.
- High specificity and good false positive control: With reversible adapter modification, the enzyme activity is blocked at room temperature, and there are no peaks within 60 minutes of LAMP/RT-LAMP.
- Excellent stability and lyophilization capability: The entire series exhibits good thermal stability and freeze-thaw stability, and the glycerol-free version facilitates the development of lyophilized reagents.
- Triton-Free and DTT-Free.
- Stable large-scale production with consistent batch-to-batch performance: Customers can confidently scale up their products for market launch.
What is the user feedback, and do you have any data to demonstrate it?
For this product upgrade, we conducted in-depth communication with several major customers who have extensive experience in using mature products and projects in multiple application fields. We benchmarked their current imported raw material performance and product upgrade requirements and proposed a new set of higher technical requirements based on the collected information.
After nearly two years of intensive development and intensive multidimensional evaluations conducted in collaboration with seed customers over the past six months, the product has been tested and approved by multiple seed customers and is being used stably. After careful evaluation from various aspects, we decided to promote and publicize it at this stage. Below are the data from user evaluations and comparative evaluations.
Partial demonstration of new BST evaluation data:
4.1 Fast amplification speed.
- A) At three concentration gradients, WS Bst DNA polymerase V2 can achieve full detection, while competitor company N shows no detection at 0.5 cp/μL.
- B) At the same concentrations (1 cp/μL and 5 cp/μL), WS Bst DNA polymerase V2 has a faster peak time (shorter Time to Result, TTR).
- C) Both WS Bst DNA polymerase V2 and competitor company N’s Bst polymerase can prevent false positives throughout the entire experimental process.
4.2 Sample compatibility
DNA system (LAMP)
- Bst DNA polymerase V2 has been tested internally and is compatible with DNA and RNA systems, providing good amplification results.
4.3 False positive control in LAMP/RT-LAMP
Reversible adapter blocking modification: Similar to antibody-based methodologies, adapter modification is also a method of enzyme activity blocking. Enzyme inactivation is achieved by the tight binding between the adapter and the polymerase at lower temperatures. At the working temperature, the binding between the adapter and the enzyme is released, initiating enzyme activity.
Example: The NTC peak chart during amplification testing using SARS-CoV-2, as shown in the figure.
Comparison of enzyme activity with Company N at 30°C and 65°C.
- A) After adapter modification and blocking, WS Bst DNA polymerase V2 (HMD7006 and HMD7007) exhibits lower enzyme activity than Company N at 30°C, resulting in higher blocking efficiency and better control of non-specific amplification.
- B) At the working temperature (65°C), it achieves complete activity release and shows no difference compared to Company N.
- C) In real-time RT-LAMP usage, no peak is observed in the No Template Control (NTC) within 60 minutes.
4.4 Inhibition resistance
Potassium Chloride (KCL)
- A) Guanidine hydrochloride and guanidine isothiocyanate are common nucleic acid extraction residues. Bst DNA polymerase V2 (HMD7005) exhibits higher relative activity than the competitor Bst under the same inhibitory concentration.
- B) Under the inhibition of KCl and NaCl salt ions, Bst DNA polymerase V2 performs better than the competitor under the same conditions.
- C) Other inhibitors, such as whole blood (including single-component), EDTA, sodium citrate, and nasopharyngeal swabs, show comparable or slightly better inhibition performance compared to the competitor.
- D) Data indicates a stronger adaptability for direct amplification from samples, facilitating the development of subsequent LAMP/RT-LAMP reagents.
4.5. Excellent stability of the lyophilized enzyme, with 100% glycerol-free.
- Both Bst DNA polymerase V2 and WS Bst DNA polymerase V2 product series maintain the same activity after incubation at 37°C for 0 days, 3 days, and 7 days.
- In order to facilitate the development of lyophilized LAMP/RT-LAMP reagents, the glycerol-free Bst DNA polymerase V2 series (HMD7005, HMD7006, HMD7007) maintains its activity after 40 repeated freeze-thaw cycles.
How to choose the product model and series?
Bst DNA polymerase V2
8U/uL，Bst DNA polymerase V2
Bst DNA polymeraseV2(Glycerol-free)
8U/uL，Bst DNA polymeraseV2(Glycerol-free)
Ws Bst DNA polymeraseV2(Glycerol-free)
8U/uL，Ws Bst DNA polymeraseV2(Glycerol-free)
WS Bst DNA polymeraseV2(High concentrationglycerol-free)
32U/uL，WS Bst DNA polymeraseV2(High concentrationglycerol-free)