食品与发酵工业 >

2020 , Vol. 46 >Issue 24: 265 - 270

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.024735

综述与专题评论

重组酶等温扩增技术在分析检测中的应用研究进展

  • 孙晓红 ,
  • 后来旺 ,
  • 李达容 ,
  • 赵勇 ,
  • 黄新新 ,
  • 何宇平 ,
  • 蓝蔚青
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  • 1(上海海洋大学 食品学院,上海, 201306)
    2(农业部水产品贮藏保鲜质量安全风险评估实验室,上海, 201306)
    3(上海水产品加工及贮藏工程技术研究中心,上海, 210306)
    4(上海海关动植物与食品检验检疫技术中心,上海, 200135)
博士,教授(蓝蔚青高级工程师为通讯作者,E-mail:wqlan@shou.edu.cn)

收稿日期: 2020-06-12

  修回日期: 2020-09-01

  网络出版日期: 2021-01-13

基金资助

2019年上海市科技兴农重点攻关项目(2019-02-08-00-10-F01149)

收起

Research progress on the application of isothermal recombinase amplification in analytical detection

  • SUN Xiaohong ,
  • HOU Laiwang ,
  • LI Darong ,
  • ZHAO Yong ,
  • HUANG Xinxin ,
  • HE Yuping ,
  • LAN Weiqing
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  • 1(College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, China)
    2(Laboratory of Quality & Safety Risk Assessment for Aquatic Products on Storage and Preservation, Ministry of Agriculture, Shanghai 201306, China)
    3(Shanghai Aquatic Products Processing and Storage Engineering Technology Research Center, Shanghai 201306, China)
    4(Technical center for animal, plant and food inspection and quarantine, Shanghai Customs District, Shanghai 200135, China)

Received date: 2020-06-12

  Revised date: 2020-09-01

  Online published: 2021-01-13

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摘要

重组酶聚合酶扩增(recombinase polymerase amplification, RPA)与重组酶介导扩增(recombinase aided amplification, RAA)是新型核酸等温扩增技术中极具代表性和前沿性的2项技术。RPA与RAA技术均可在常温下进行等温扩增,还可在简单设备甚至低资源环境下实现快速扩增,具有特异性强、灵敏度高、操作简便、检测时间短等优点。该文在简述RPA与RAA技术扩增原理的基础上,重点阐述了主要针对重组酶等温扩增技术在分析检测中的应用研究进展,提出存在问题与解决办法,并对其未来发展前景予以展望,以期为拓展2项技术的应用领域提供理论参考。

关键词： 重组酶聚合酶扩增; 重组酶介导扩增; 分析检测; 研究进展

本文引用格式

孙晓红 , 后来旺 , 李达容 , 赵勇 , 黄新新 , 何宇平 , 蓝蔚青 . 重组酶等温扩增技术在分析检测中的应用研究进展[J]. 食品与发酵工业, 2020 , 46(24) : 265 -270 . DOI: 10.13995/j.cnki.11-1802/ts.024735

Abstract

Recombinase polymerase amplification (RPA) and recombinase aided amplification (RAA) are two representative and frontier technologies in the novel nucleic acid isothermal amplification. Both RPA and RAA can be used for isothermal amplification at room temperature and for rapid amplification in simple equipment or even in resource-limited environment. They have the advantages of high specificity, high sensitivity, simple operation and short detection time. Based on the brief introduction of the amplification principle of RPA and RAA, this paper focused on the application and progress of isothermal recombinase amplification assay in analysis and detection. The problems and solutions were put forward and the development of isothermal recombinase amplification was prospected, which can provide theoretical reference for expanding the application field of RPA and RAA in the future.

Key words： recombinase polymerase amplification(RPA); recombinase aided amplification(RAA); analytical detection; research progress

参考文献

[1] 李聪,刘健慧,李志辉, 等.4种食源性致病菌的多重PCR快速检测方法研究[J].食品研究与开发, 2019, 40(21):164-169.
LI C, LIU J H, LI Z H, et al.Study on multiplex PCR of rapid detection method for four foodborne pathogens[J].Food Research and Development, 2019, 40(21):164-169.
[2] 崔丙健,高峰,胡超, 等.不同再生水灌溉方式对土壤-辣椒系统中细菌群落多样性及病原菌丰度的影响[J].环境科学, 2019, 40(11):5 151-5 163.
CUI B J, GAO F, HU C, et al.Effect of different reclaimed water irrigation methods on bacterial community diversity and pathogen abundance in the soil-pepper ecosystem[J].Environmental Science, 2019, 40(11):5 151-5 163.
[3] PIEPENBURG O, WILLIAMS C H, STEMPLE D L, et al.DNA detection using recombination proteins[J].Plos Biology, 2006, 4(7):e204.
[4] LÜ B, CHENG H R, YAN Q F, et al.Recombinase-aid amplification: A novel technology of in vitro rapid nucleic acid amplification[J].Scientia Sinica Vitae, 2010, 40:983-988.
[5] KUZMINOV A.DNA replication meets genetic exchange:Chromosomal damage and its repair by homologous recombination[J].Proceedings of the National Academy of Sciences of the United States of America, 2001, 98(15):8 461-8 468.
[6] WANG Z, YANG P P, ZHANG Y H, et al.Development of a reverse transcription recombinase polymerase amplification combined with lateral-flow dipstick assay for avian influenza H9N2 HA gene detection[J].Transboundary and Emerging Diseases, 2018, 66(1):546-551.
[7] 鲍金圭,李崇建,江慧,等.2016-2018年钦州市手足口病住院患儿病原学检测结果分析[J].检验医学与临床, 2019, 16(24):3 592-3 595.
BAO J G, LI C J, JIANG H, et al.The pathogenic composition of hospitalized children with hand, foot and mouth disease in Qinzhou during 2016-2018[J].Laboratory Medicine and Clinic, 2019, 16(24):3 592-3 595.
[8] YIN D, ZHU Y N, WANG K F, et al.Development and evaluation of a rapid recombinase polymerase amplification assay for the detection of human enterovirus 71[J].Archives of Virology, 2018, 163(9):2 459-2 463.
[9] LI X N, SHEN X X, LI M H, et al.Applicability of duplex real time and lateral flow strip reverse-transcription recombinase aided amplification assays for the detection of Enterovirus 71 and Coxsackievirus A16[J]. Virology Journal, 2019, 16(1):166.
[10] WANG K F, WU Y, YIN D, et al.Development and evaluation of a rapid recombinase polymerase amplification assay for detection of coxsackievirus A6[J]. Archives of Virology, 2017, 162(1):287-290.
[11] YAN T F, LI X N, WANG L, et al.Development of a reverse transcription recombinase-aided amplification assay for the detection of coxsackievirus A10 and coxsackievirus A6 RNA[J].Archives of Virology, 2018, 163(6):1 455-1 461.
[12] SHEN X X, QIU F Z, SHEN L P, et al.A rapid and sensitive recombinase aided amplification assay to detect hepatitis B virus without DNA extraction[J].BMC Infectious Diseases, 2019, 19(1):229.
[13] BAI X D, MA X J, LI M H, et al.Field applicable detection of hepatitis B virus using internal controlled duplex recombinase-aided amplification assay and lateral flow dipstick assay[J].Journal of Medical Virology, 2020,92(12);3 344-3 353.
[14] QI J J, LI X N, ZHANG Y, et al.Development of a duplex reverse transcription recombinase-aided amplification assay for respiratory syncytial virus incorporating an internal control[J].Archives of Virology, 2019, 164(7):1 843-1 850.
[15] XI Y, XU C Z, XIE Z Z, et al.Development of a reverse transcription recombinase polymerase amplification assay for rapid detection of human respiratory syncytial virus[J].Molecular and Cellular Probes, 2019, 45:8-13.
[16] 孙晓红,后来旺,郜蓉.一种重组酶等温扩增技术检测副溶血性弧菌的方法:中国, CN 110564881A [P].2019-10-21.
SUN X H, HOU L W, GAO R.Method for detecting Vibrio parahaemolyticus by isothermal recombinase aided amplification:China, CN 110564881A [P].2 019-10-21.
[17] 魏莹,郭利川,张小平,等.重组酶介导扩增方法快速检测A族乙型溶血性链球菌[J].中国国境卫生检疫杂志, 2018, 41(5):314-316;323.
WEI Y, GUO L C, ZHANG X P, et al.Establishment of recombinase aided amplification assay for group A streptococcus pyogens detection[J].Chinese Journal of Frontier Health and Quarantine, 2018, 41(5):314-316;323.
[18] DU X J, ZANG Y X, LIU H B, et al.Recombinase polymerase amplification combined with lateral flow strip for Listeria monocytogenes detection in food[J].Journal of Food Science, 2018, 83(4):1 041-1 047.
[19] 周广彪,段建发,胡晓珊, 等.重组酶聚合酶扩增技术(RPA)检测拟态弧菌[J].检验检疫学刊, 2018, 28(5):1-4;16.
ZHOU G B, DUAN J F, HU X S, et al.Detection of Vibrio Mimicus heat-labile hemolysin by recombinase polymerase amplification[J].Journal of Inspection and Quarantine, 2018, 28(5):1-4;16.
[20] LI S Z, ZHENG H, GAO J, et al.Endemic status of schistosomiasis in the People’s Republic of China in 2012[J].Chinese Journal of Schistosomiasis Control, 2013, 25(6):557-563.
[21] BERGQUIST R, JOHANSEN M V, UTZINGER J.Diagnostic dilemmas in helminthology:What tools to use and when? [J].Trends in Parasitology, 2009, 25(4):151-156.
[22] 银娜,梁俊,高志贤.银纳米簇用于食品包装污染物的检测及其抗菌性能的应用[J].包装工程, 2019, 40(21):44-50.
YIN N, LIANG J, GAO Z X.Silver nanoclusters for the detection of food packaging contaminants and their application in antibacterial properties[J].Packaging Engineering, 2019, 40(21):44-50.
[23] SONG Z, TING L, KUN Y, et al.Establishment of a recombinase-aided isothermal amplification technique to detect Schistosoma japonicum specific gene fragments[J].Chinese Journal of Schistosomiasis Control, 2018, 30(3):273-277.
[24] SUN K, XING W, YU X, et al.Recombinase polymerase amplification combined with a lateral flow dipstick for rapid and visual detection of Schistosoma japonicum[J].Parasite Vectors, 2016, 9(1):476.
[25] 唐慧骥,党志浩,郭长宁,等.应用重组酶介导核酸扩增技术快速鉴定新菠萝灰粉蚧[J].植物检疫, 2019, 33(2):37-42.
TANG H J, DANG Z H, GUO C N, et al.Rapid detection for Dysmicoccus neobrevipes by recombinase-aid amplification[J].Plant Quarantine, 2016, 9(1):476.
[26] 周鸿让, 陈木新, 余晴, 等.重组酶介导的等温核酸扩增技术检测多房棘球绦虫方法的建立及初步应用[J].中国血吸虫病防治杂志, 2020, 32(2):168-173;180.
ZHOU H R, CHEN M X, YU Q, et al.Establishment of a recombinase-aided isothermal amplification assay for nucleic acid detection of Echinococcus multilocularis and its preliminary application[J].Chinese Journal of Schistosomiasis Control, 2020, 32(2):168-173;180.
[27] 周鸿让,茅光耀,王晓玲,等.重组酶介导的多重核酸等温扩增法鉴别细粒棘球绦虫G1和多房棘球绦虫技术的建立与应用[J].中国寄生虫学与寄生虫病杂志, 2020,38(3):310-316.
ZHOU H R, MAO G Y, WANG X L, et al.Establishment and application of a multiplex recombinase-aided isothermal amplification technique for identifying Echinococcus granulosus G1 and Echinococcus multilocularis[J].Chinese Journal of Parasitology and Parasitic Diseases, 2020,38(3):310-316.
[28] 佚名. 温州警方查获制售假牛肉干案一年销售达百万元[J].中国防伪报道, 2017(5):85-86.
YI M.Police in Wenzhou have uncovered a case in which fake beef jerky was sold for up to one million yuan a year[J].China Anti-Counterfeiting Report, 2017(5):85-86.
[29] 韩利. 牛丸无牛肉虾丸中掺鸭 成都一公司掺假掺杂被罚40万[J].广西质量监督导报, 2016(12):35-36.
HAN L.A company in Chengdu was fined 400,000 yuan for adulterating beef and shrimp balls[J].Guangxi Quality Supervision Guide Periodical, 2016(12):35-36.
[30] 王耕,张薇,陈安东,等.肉类食品的动物物种基因鉴别及等温扩增荧光(IAFT)快速检测技术的建立[J].生物化工, 2016, 5(2):21-25.
WANG G, ZHANG W, CHEN A D, et al.Identification of animal species in meat food and establishment of rapid detection technology for isothermal amplification of fluorescence (IAFT) [J].Biological Chemical Engineering, 2016, 5(2):21-25.
[31] 肖蕾,蓝蔚青,孙晓红,等.金枪鱼常用保鲜方式及品质检测技术研究进展[J].包装工程, 2017, 38(5):115-120.
XIAO L, LAN W Q, SUN X H, et al.Research progress in commonly used preservation methods and quality detection techniques of Tuna[J].Packaging Engineering, 2017, 38(5):115-120.
[32] SUN X H, XU Q, PAN Y J, et al.A loop-mediated isothermal amplification method for rapid detection of Vibrio parahaemolyticus in seafood[J].Annals of Microbiology, 2012, 62(1):263-271.
[33] 郭燕华,王德莲,王强,等.重组酶介导等温扩增技术快速检测牛肉及牛肉制品中的牛源性成分[J].食品安全质量检测学报, 2017, 8(5):1 745-1 749.
GUO Y H, WANG D L, WANG Q, et al.Determination of bovine ingredient in beef and its derivates with recombinase polymerase mediated isothermal amplification[J].Journal of Food Safety & Quality, 2017, 8(5):1 745-1 749.
[34] 林霖,冯荣虎,王坤,等.牛肉真伪鉴别荧光RPA现场快检方法建立[J].食品科技, 2018, 43(6):322-326.
LIN L, FENG R H, WANG K, et al.Construct beef authentication on-site rapid detection method based on fluorescent RPA technology[J].Food Science and Technology, 2018, 43(6):322-326.
[35] 刘静,武国干,吴潇,等.重组酶聚合酶扩增技术快速检测转基因玉米Bt11[J].上海农业学报, 2018, 34(1):20-24.
LIU J, WU G G, WU X, et al.Rapid detection of genetically modified maize Bt11 by recombinase polymerase amplification assay[J].Acta Agriculturae Shanghai, 2018, 34(1):20-24.
[36] 谢实龙. 转基因成分重组酶聚合酶扩增(RPA)快速检测方法的建立[D].阜阳:阜阳师范学院, 2019.
XIE S L.The application of recombinase polymerase amplification technology in transgenic detection [D].Fuyang:Fuyang Normal University, 2019.
[37] 廖新辉,易浔飞,崔智慧,等.用重组酶介导等温核酸扩增技术检测烟曲霉[J].临床检验杂志, 2015, 33(3):214-218.
LIAO X H, YI X F, CUI Z H, et al.Detection for Aspergillus fumigatus by recombinase polymerase amplification assay[J].Chinese Journal of Clinical Laboratory Science, 2015, 33(3):214-218.
[38] 刘爽,黄广涛,龚雅利, 等.基于重组酶聚合酶扩增技术建立实时荧光法快速检测鲍曼不动杆菌的研究[J].中国病原生物学杂志, 2019, 14(3):311-314.
LIU S, HUANG G T, GONG Y L, et al.Rapid detection of Acinetobacter baumannii by real-time fluorescence method based on recombinase polymerase amplification[J].Journal of Pathogen Biology, 2019, 14(3):311-314.
[39] 黄纪徽,吴山楠,王丹云, 等.虾肝肠胞虫实时荧光RAA快速检测方法的建立[J].养殖与饲料, 2018(11):13-16.
HUANG J H, WU S N, WANG D Y, et al.Establishment of a fast real-time recombinase aided amplification detection method for shrimp Hepatocellular cytospora[J].Animals Breeding and Feed, 2018(11):13-16.
[40] MA B, FANG J, LIU W, et al.A simple and efficient method for potential point-of-care diagnosis of human papillomavirus genotypes:Combination of isothermal recombinase polymerase amplification with lateral flow dipstick and reverse dot blot[J].Analytical and Bioanalytical Chemistry, 2019, 411(28):7 451-7 460.
[41] CHEN J S, MA E, HARRINGTON L B, et al.CRISPR-Cas12a target binding unleashes indiscriminate single-stranded DNase activity[J].Science, 2018, 360(6 387):436-439.
[42] LIU H B, ZANG Y X, DU X J, et al.Development of an isothermal amplification-based assay for the rapid visual detection of Salmonella bacteria[J].Journal of Dairy Science, 2017, 100(9):7 016-7 025.
[43] 王亚磊,张海洋,王权,等.产单核细胞李氏杆菌RPA-LF快速检测方法的建立[J].中国兽医科学, 2020, 50(1):1-9.
WANG Y L, ZHANG H Y, WANG Q, et al.Rapid detection of Listeria monocytogenes with isothermal recombinase polymerase amplification and lateral flow analysis[J].Chinese Veterinary Science, 2020, 50(1):1-9.
[44] HIGGINS M, RAVENHALL M, WARD D, et al.PrimedRPA:Primer design for recombinase polymerase amplification assays[J].Bioinformatics, 2019, 35(4):682-684.
[45] MOHAMMAD U, TEIXEIRA DA SILVA J, Zou Y Y, et al.Successful amplification of DNA using recombinase-aided amplification (RAA) and its application to seven horticultural crops[J].Biotechnology, 2016, 2(12):7.
[46] ZACHARY A C, BRITTANY R, REBECCA R K.Equipment-free incubation of recombinase polymerase amplification reactions using body heat[J].PloS One, 2014, 9(11):e112 146.
[47] 杭州优思达生物技术有限公司. 一种核酸一体化检测方法及检测试剂管:中国, CN 108796038A [P].2018-06-26.
Hangzhou Ustar Biotechnologies .The invention relates to a nucleic acid integration detection method and a detection reagent tube:China, CN 108796038A [P].2018-06-26.
[48] 李伟,庄濠宇,温尔英,等.添加扩增内标的重组酶聚合酶扩增技术(RPA-IAC)检测创伤弧菌[J].检验检疫学刊, 2019, 29(3):5-8.
LI W, ZHUANG H Y, WEN E Y, et al.Recombinant enzyme polymerase amplification (RPA-IAC) with amplified internal standard for detection of Vibrio vulnificus[J].Journal of Inspection and Quarantine, 2019, 29(3):5-8.
[49] YANG H L, WEI S, GOONERATNE R, et al.Development of a recombinase polymerase amplification assay for Vibrio parahaemolyticus detection with an internal amplification control[J].Canadian Journal of Microbiology,2018, 64(4):223-230.
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