What is RPA?
概念：重組酶聚合酶擴(kuò)增(Recombinase Polymerase Amplification，RPA)，被稱為是可以替代PCR的核酸檢測(cè)技術(shù)。RPA技術(shù)主要依賴于三種酶：能結(jié)合單鏈核酸(寡核苷酸引物)的重組酶、單鏈DNA結(jié)合蛋白(SSB)和鏈置換DNA聚合酶。這三種酶的混合物在常溫下也有活性，最佳反應(yīng)溫度在37°C左右。
RPA技術(shù)原理：
重組酶與引物結(jié)合形成的蛋白-DNA復(fù)合物，能在雙鏈DNA中尋找同源序列。一旦引物定位了同源序列，就會(huì)發(fā)生鏈交換反應(yīng)形成并啟動(dòng)DNA合成，對(duì)模板上的目標(biāo)區(qū)域進(jìn)行指數(shù)式擴(kuò)增。被替換的DNA鏈與SSB結(jié)合，防止進(jìn)一步替換。在這個(gè)體系中，由兩個(gè)相對(duì)的引物起始一個(gè)合成事件。整個(gè)過(guò)程進(jìn)行得非常快，一般可在十分鐘之內(nèi)獲得可檢出水平的擴(kuò)增產(chǎn)物。
Who uses RPA?
以此為基礎(chǔ)的TwistAmp? 核酸擴(kuò)增產(chǎn)品，能夠在15分鐘內(nèi)進(jìn)行常溫下的單分子核酸檢測(cè)該技術(shù)對(duì)硬件設(shè)備的要求很低，特別適合用于體外診斷、獸醫(yī)、食品安全、生物安全、農(nóng)業(yè)等領(lǐng)域。
RPA等溫?cái)U(kuò)增試劑盒配套試紙條發(fā)表的文獻(xiàn)
1、Piepenburg O., Williams C.H., Stemple D.L., Armes N.A.
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9、Rohrman B. et al., Inhibiotion of Recombinase Polymerase Amplification by Background DNA: A Lateral Flow-Based Method for Enriching Target DNA. Anal. Chem. 201587 (3), 1963-1967http://pubs.acs.org/doi/pdf/10.1021/ac504365v
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11、Zhao G. et al., Rapid visual detection of Mycobacterium avium subsp.paratuberculosis by recombinase polymerase amplification combined with a lateral flow dipstick. J Vet Sci. 2018 Mar 31;19(2):242-250.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5879072/pdf/jvs-19-242.pdf
12、Qi Y. et al., Development of a rapid and visual nucleotide detection method for a Chinese epidemicstrain of Orientia tsutsugamushi based on recombinase polymerase amplification assay and lateral flow test. Int J Infect Dis. 2018 May;70:42-50.https://www.ijidonline.com/article/S1201-9712(18)30062-6/pdf
13、Ahmed F. A. et al., Genome-informed diagnostics for specific and rapid detection ofPectobacterium species using recombinase polymerase amplification coupled with a lateral flow device.Sci Rep. 2018 Oct 29;8(1):15972
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14、Qi Y. et al., Development of a rapid and visual detection method for Rickettsia rickettsii combining recombinase polymerase assay with lateral flow test. PLoS One. 2018Nov 26;13(11)https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6257923/pdf/pone.0207811.pdf
15、Wang H. et al., Rapid visual detection of cyprinid herpesvirus 2 by recombinase polymeraseamplification combined with a lateral flow dipstick. J Fish Dis. 2018May 28https://onlinelibrary.wiley.com/doi/abs/10.1111/jfd.12808
16、Wang H. et al., Development and evaluation of serotype-specific recombinase polymeraseamplification combined with lateral flow dipstick assays for the diagnosis of foot-and-mouthdisease virus serotype A, O and Asia1. BMC Vet Res. 2018Nov 20;14(1)https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6245561/pdf/12917_2018_Article_1644.pdf
17、Soliman H. et al., Recombinase polymerase amplification assay combined with a lateral flowdipstick for rapid detection of Tetracapsuloides bryosalmonae, the causative agent of proliferativekidney disease in salmonids. Parasit Vectors. 2018 Apr11;11(1):234.https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5896054/pdf/13071_2018_Article_2825.pdf
18、Soliman H. and El-Matoubli M., Rapid detection and differentiation ofcarp oedema virus and cyprinid herpes virus-3 in koi and common carp. J Fish Dis. 2018 May;41(5):761-772.
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https://www.sciencedirect.com/science/article/pii/S0003269718307085?via%3Dihub
20、Zhao G. et al., Use of a recombinase polymerase amplification commercial kit for rapid visual detection of Pasteurella multocida. BMC Vet. Res. 2019; 15:154
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1475771/pdf/pbio.0040204.pdf
21、Li T.-T. et al., Rapid and Visual Detection of Trichinella Spp. Using a Lateral Flow Strip-Based Recombinase Polymerase Amplification (LF-RPA) Assay. Front. Cell. Infect. Microbiol.2019; 9 1.https://www.frontiersin.org/articles/10.3389/fcimb.2019.00001/full
22、Ma Q. et al., Development of a lateral flow recombinase polymerase amplification assay for rapid and visual detection of Cryptococcus neoformans/C. gattii in cerebral spinal fluid. BMC Infect. Dis.2019; 19:108
https://www.researchgate.net/publication/330863615_Development_of_a_lateral_flow_recombinase_polymerase_amplification_assay_for_rapid_and_visual_detection_of_Cryptococcus_neoformansC_gattii_in_cerebral_spinal_fluid/fulltext/5c59049792851c22a3aa5082/330863615_Development_of_ a_lateral_flow_recombinase_polymerase_amplification_assay_for_rapid_and_visual_detection_of_ Cryptococcus_neoformansC_gattii_in_cerebral_spinal_fluid.pdf?origin=publication_detail
23、Peng Y. et al., Rapid detection of Burkholderia pseudomallei with a lateral flow recombinasepolymerase amplification assay. PLoSONE 2019; 14(7)
https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0213416&type=printable
24、Sullivan, T. J., Dhar, A. K., Cruz-Flores, R., & Bodnar, A. G. (2019). Rapid, CRISPR-Based, Field-Deployable Detection Of White Spot Syndrome Virus In Shrimp. Scientific Reports, 9(1), 1–7. https://doi.org/10.1038/s41598-019-56170-y
默瑞(上海)生物科技有限公司，作為專(zhuān)業(yè)的生命科學(xué)產(chǎn)品推廣與服務(wù)商，以“創(chuàng)新，專(zhuān)業(yè)，共贏”的理念，不斷強(qiáng)化服務(wù)體系，提升客戶體驗(yàn)，以莫納生物產(chǎn)品為主線，結(jié)合國(guó)內(nèi)外優(yōu)質(zhì)產(chǎn)品提供高標(biāo)準(zhǔn)完整解決方案，幫助科研客戶和生物技術(shù)企業(yè)高效創(chuàng)新，加速轉(zhuǎn)化，創(chuàng)造更多優(yōu)質(zhì)產(chǎn)品，提升社會(huì)福祉，實(shí)現(xiàn)多方共贏。
默瑞生物創(chuàng)立以來(lái)，已經(jīng)陸續(xù)導(dǎo)入TwistDx，enzymatics，KAPA BIOSYSTEMS，BioDynami，Jackson，Echelon，ChromoTek，Nanocs，Biotechrabbit，Lee Biosolutions, magtivio等優(yōu)質(zhì)國(guó)際品牌，逐步培育成為專(zhuān)業(yè)用戶的首選。
