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客戶服務熱線
400-0470-600
微信
13903605303
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| HaiGene Bst4.2 DNA聚合酶系列LAMP相關試劑是HaiGene精心研制的高性能的LAMP擴增用試劑，通用于DNA或RNA的靶標檢測。在快速、高耐受性的基礎上，不僅實現穩定、快速、高靈敏度的擴增性能，在HotStart性能的加持下，還實現了假陽性的大幅下降。 制品性能特點： （1） Bst4.0全系包含熱啟動Aptamer，該配體確保酶在<30℃時，酶活封閉效率>90%,在>60℃時1min內完全釋放酶活。該特性利于室溫建立反應體系，并大幅降低了低溫條件下的非特異擴增。 （2） Bst4.2系列反應溫度提升到70℃，大幅降低引物Dimer的形成，提高擴增特異性，并使得粗樣品核酸釋放更加充分。 （3） Bst4.2系列包含Helicaser，因此，允許在不使用F3/B3引物的情況下進行eLAMP擴增（easy LAMP），并允許FIP/BIP的引物用量降低一倍。這將進一步降低非特異擴增，并使得擴增均一性大幅提升。 （4） 預凍干液體形式的試劑，賦予多用途性，不僅能滿足常規的科研需求，還使得凍干產品開發和生產在一種試劑下同步完成。 |
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性能展示 |
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| 1、Bst 4.2 HotStart功能展示 采用熒光酶活測定法在30℃和60℃條件下測定HotStart Bst 4.2，結果顯示與未加入Aptamer的對照比較來看。HotStart Bst 4.2在30℃條件下酶活幾乎無活性，而60℃條件下1min內酶活完全釋放，恢復100%活性。 |
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| 2、以Bst 4.2 SYBR Green試劑進行標準LAMP和eLAMP擴增比較 以human Total RNA為模板，采用RnaseP LAMP Primer進行標準LAMP和eLAMP 擴增。在25ul擴增體系中加入10ng、1ng和0ng的RNA，70℃反應40min。從擴增結果可獲得，在Helicaser的加持下F3/B3可完全去除，對擴增速度幾乎無影響，并且非特異性擴增得到改善。 6 Primer LAMP引物濃度：FIP/BIP=0.8 μM; LoopF/B=0.4 μM; F3/B3=0.1 μM。 4 Primer eLAMP引物濃度：FIP/BIP=0.8 μM; LoopF/B=0.4 μM。 |
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| 3、以Bst 4.2 Basic試劑進行DP-eLAMP探針法擴增 在眾多的探針法LAMP測試中，DP-LAMP探針法(Displaceable Probe LAMP)為HaiGene推薦的使用方法（參考PMID: 33626039）。在LAMP擴增中將LF或LB引物退火至互補的猝滅oligo，利用鏈置換活性獲得游離的熒光信號（可參考HaiGene A3831-57說明書資料）。該方案可進一步降低LAMP的非特異擴增。下圖為DP-eLAMP進行三種熒光標記探針的實驗數據。結果顯示HotStart Bst4.2總能獲得高特異性和高重復性的擴增 |
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| 4、以Bst4.2 L-HNB 試劑進行L-HNB (紫羅蘭-淡綠) 變色反應（肉眼觀察） 以2019-nCoV體外轉錄RNA為模板，采用eLAMP擴增法（FIP/BIP=0.8 μM; LoopF/B=0.4 μM）, 對10000、1000、100、25copies/管的陽性RNA進行檢測，ddH20為陰性對照（4重復）。下圖為70℃反應30min 后的檢測結果。結果表明L-HNB法獲得清晰易于區分的顏色變化，極易區分陽性和陰性擴增。 |
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| 5、以Bst4.2 L-HNB (紫羅蘭-淡綠) 變色反應對反應緩沖鹽Tris的耐受性測試 以2019-nCoV體外轉錄RNA為模板，采用eLAMP擴增法（FIP/BIP=0.8 μM; LoopF/B=0.4 μM）, 對1000copies/管的陽性RNA進行檢測（2重復），ddH20為陰性對照（2重復）。下圖為70℃反應30min 后的檢測結果。結果表明L-HNB法，在1x濃度下可耐受10mM Tris-HCl的緩沖鹽，陽性樣本仍然可以充分的變色。該變色方法不受樣本中緩沖鹽影響。 |
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應用LAMP產品文獻 |
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| 1. Rapid Visual Detection of Elite Erect Panicle Dense and Erect Panicle 1 Allele for Marker-Assisted Improvement in Rice (Oryza sativa L.) Using the Loop-Mediated Isothermal Amplification Method. Curr Issues Mol Biol. 2024 Jan 4;46(1):498–512. 2. Rapid Visual Detection of High Nitrogen-Use Efficiency Gene OsGRF4 in Rice (Oryza sativa L.) Using Loop-Mediated Isothermal Amplification Method. Genes (Basel). 2023 Sep 23;14(10):1850. 3. Rapid and visual identification of HIV-1 using reverse transcription loop-mediated isothermal amplification integrated with a gold nanoparticle-based lateral flow assay platform. Front Microbiol. 2023 Jul 12;14:1230533. 4. Rapid Detection of Streptococcus mutans Using an Integrated Microfluidic System with Loop-Mediated Isothermal Amplification. J Microbiol Biotechnol. 2023 May 19;33(8):1101–1110. 5. Rapid point-of-care detection of BK virus in urine by an HFman probe-based loop-mediated isothermal amplification assay and a finger-driven microfluidic chip.PeerJ. 2023 Mar 8;11:e14943. 6. Rapid detection of monkeypox virus and monkey B virus by a multiplex loop-mediated isothermal amplification assay. J Infect. 2023 Feb 13;86(4):e114–e116. 7. SMART: A Swing-Assisted Multiplexed Analyzer for Point-of-Care Respiratory Tract Infection Testing. Biosensors (Basel). 2023 Feb 4;13(2):228. 8. An HFman Probe-Based Multiplex Reverse Transcription Loop-Mediated Isothermal Amplification Assay for Simultaneous Detection of Hantaan and Seoul Viruses. Diagnostics (Basel). 2022 Aug 10;12(8):1925. 9. Real-Time Detection of LAMP Products of African Swine Fever Virus Using Fluorescence and Surface Plasmon Resonance Method. Biosensors (Basel). 2022 Apr 3;12(4):213. 10.Multiplex, Real-Time, Point-of-care RT-LAMP for SARS-CoV-2 Detection Using the HFman Probe.ACS Sens. 2022 Feb 22;7(3):730–739. 11.Deep‐dLAMP: Deep Learning‐Enabled Polydisperse Emulsion‐Based Digital Loop‐Mediated Isothermal Amplification. Adv Sci (Weinh). 2022 Jan 24;9(9):2105450. 12.Development of a TaqMan loop-mediated isothermal amplification assay for the rapid detection of pigeon paramyxovirus type 1. Arch Virol. 2021 Mar 23;166(6):1599–1605. 13.Rapid, visual, label-based biosensor platform for identification of hepatitis C virus in clinical applications. BMC Microbiol. 2024 Feb 28;24:68. 14.A novel extraction-free dual HiFi-LAMP assay for detection of methicillin-sensitive and methicillin-resistant Staphylococcus aureus.Microbiol Spectr. 2024 Feb 20;12(4):e04133-23. 15.A duplex HiFi-LAMP assay for screening of two novel human circoviruses HCirV-1 and HCirV-2. BMC Infect Dis. 2024 Dec 5;24:1388. 16.Point-of-care test of blood Plasmodium RNA within a Pasteur pipette using a novel isothermal amplification without nucleic acid purification. Infect Dis Poverty. 2024 Oct 31;13:80. 17.Rapid detection of human influenza A viruses by HFman probe-based loop-mediated isothermal amplification assays. Heliyon. 2023 Oct 25;9(11):e21591. |
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