环介导等温扩增技术及其应用

doi:10.3969/j.issn.1674-5817.2014.02.018

摘要/Abstract

摘要： 环介导等温扩增法(LAMP)是一种新的核酸扩增法。该方法在一定温度下一步即可完成扩增反应。由于其扩增效率极高、反应简单快速、高特异性和反应结果只需肉眼观察的特点,LAMP法已广泛应用于病毒、细菌等的定性定量检测和临床疾病的诊断。本文介绍了LAMP方法的原理及其在微生物分子检测方面的应用。

关键词: 环介导等温扩增法(LAMP), 核酸扩增, 基因检测

Abstract: Loop-mediated isothermal amplification(LAMP) is a novel nucleic acid amplification method. At a certain temperature, LAMP performed only one step to terminate the process. Due to its high efficiency, simplicity, rapidly, high specificity and the result of LAMP is easily detected by naked eyes, LAMP has been widely applied in the qualitative and quantitative detection of viruses, bacteria and clinical diagnosis of disease. In this paper, we introduced the principle of LAMP and its applicability in molecular detection and diagnosis.

Key words: Loop-mediated isothermal amplification(LAMP), Nucleic acid amplification, Gene detection

中图分类号:

Q95-33

赵丽娟,周洁,高诚. 环介导等温扩增技术及其应用[J]. 实验动物与比较医学, 2014, 34(2): 162-166. DOI: 10.3969/j.issn.1674-5817.2014.02.018.

ZHAO Lijuan,ZHOU Jie,GAO Cheng. Loop-mediated Isothermal Amplification Technique and Its Application[J]. Laboratory Animal and Comparative Medicine, 2014, 34(2): 162-166. DOI: 10.3969/j.issn.1674-5817.2014.02.018.

参考文献

[1] Notomi T, Okayama H, Masubuchi H, et al.Loop-mediated isothermal amplification of DNA[J]. Nucleic Acids Res, 2000, 28(12): 63-69.
[2] Mori Y, Nagamine K, Tomita N, et al.Detection of loop-mediated isothermal amplification reaction by turbidity derived from magnesium pyrophosphate formation[J]. Biochem Bioph Res Commun, 2001, 289(1): 150-154.
[3] 张小飞, 白杰英, 赵爽, 等. 环介导等温扩增技术的研究进展和微生物检测应用[J]. 实验动物与比较医学, 2011, 31(3): 226-230.
[4] Fang XE, Li J, Chen Q.One new method of nucleic acid amplification-loop-mediated isothermal amplification of DNA[J]. ViroSin, 2008, 23(3): 167-172.
[5] Nose K, Nagamine K, Tokuda J, et al.Polyethylene glycol accelerates loop-mediated isothermal amplification (LAMP) reaction[J]. Yakugaku Zasshi, 2013, 133(10): 1121-1126.
[6] Imai M, Ninomiya A, Minekawa H, et al.Rapid diagnosis of H5N1 avian influenza virus infection by newly developed influenza H5 hemagglutinin gene-specific loop-mediated isothermal amplification method[J]. J Virol Methods, 2007, 141(2):173-180.
[7] Dinh DT, Le MT, Vuong CD, et al.An Updated Loop-Mediated Isothermal Amplification Method for Rapid Diagnosis of H5N1 Avian Influenza Viruses[J]. Trop Med Int Health, 2011, 39(1):3-7.
[8] Zhang J, Feng Y, Hu D, et al.Rapid and sensitive detection of H7N9 avian influenza virus using reverse transcription loop-mediated isothermal amplification[J]. J Clin Microbiol, 2013, 51(11):3760-3764.
[9] 陈蕾, 范学政, 王琴, 等. 中国畜牧兽医学会2008年学术年会暨第六届全国畜牧兽医青年科技工作者学术研讨会论文集[A]. 中国畜牧兽医学会, 2008, 209-213.
[10] Yin SH, Shang YJ, Zhou GQ, et al.Development and evaluation of rapid detection of classical swine fever virus by reverse transcription loop-mediated isothermal amplification(RT-LAMP)[J]. Biotechnology, 2010, 146(4): 147-150.
[11] Miren Iturriza-Gómara, Jaqueline Xerry, Chris I Gallimore, Evaluation of the Loopamp (loop-mediated isothermal amplification) kit for detecting Norovirus RNA in faecal samples[J]. J Clin Virol, 2008, 42(4):389-393.
[12] Li Qiming, Ma Xuejun, Zhou Rui, et al.Detection of HCV gene by reverse transcription loop-mediated isothermal amplification method[J]. Chin J Virol, 2006, 22(5): 39-44.
[13] Qiong Li, Zhiqin Yue, Hong Liu, et al.Development and evaluation of a loop-mediated isothermal amplification assay for rapid detection of lymphocystis disease virus[J]. J Virol Methods, 2010, 163(2):378-384.
[14] 黄鹤, 李应国, 肖进文, 等. 羊痘病毒环介导等温扩增(LAMP)检测方法的建立[J]. 中国畜牧兽医, 2012, 39(6): 72-75.
[15] Qiao J, Wang J, Meng Q, et al.Rapid detection of Akabane virus by a novel reverse transcription loop-mediated isothermal amplification assay (RT-LAMP). Virol J, 2013, 10: 288.
[16] Shen W, Tuo D, Yan P, et al.Detection of Papaya leaf distortion mosaic virus by reverse-transcription loop-mediated isothermal amplification[J]. J Virol Methods, 2013, 195: 174-179.
[17] Dukes JP, King DP, Alexandefsen S.Novel reverse transcription loop-mediated isothermal amplification for rapid detection of foot-and-mouth disease virus[J]. Arch Virol, 2006, 151(6): 1093-1106.
[18] Maruyama F, KenzakaT, Yamaguchi N, et al. Detection of bacteria carrying the stx2 gene by in situ loop-mediated isothermal amplication[J]. Appl Environ Microbiol, 2003, 69(8): 5023-5028.
[20] 吕恒, 张锦海, 陈凤娟, 等. 大肠埃希菌O157:H7环介导等温扩增可视化检测方法的建立[J]. 中国病原生物学杂志, 2013, 8(4): 289-292.
[18] 朱杰仪, 谭实勇, 曾小娜, 等. 副猪嗜血杆菌环介导等温扩增检测方法的建立[J]. 中国畜牧兽医, 2010, 37(2):204-206.
[21] Mitarai S, Okumura M, Toyota E, et al.Evaluation of a simple loop-mediated isothermal amplifi cation test kit for the diagnosis of tuberculosis[J]. Int J Tuberc Lung Dis, 2011, 15(9):1211-1217.
[22] Goto M, Hayashidani H, Takatori K, et al.Rapid detection of enterotoxigenic Staphylococus aureus harbouring genes for four classical enterotoxins,SEA,SEB,SECandSED,by loop-mediated isothermal amplification assay[J]. Lett Appl Microbiol, 2007, 45(1):100-107.
[23] Han FF, Ge BL.Evaluation of a loop-mediated isothermal amplification assay for detecting Vibrio vulnificus in raw oysters[J]. Foodborne Pathog Dis, 2008, 5(3): 311-320.
[24] 魏东, 汪洁英, 王国治. 鼠疫耶尔森氏菌环介导等温扩增检测方法的建立[J]. 中国卫生检验杂志, 2012, 22(9):2074-2077.
[25] Yang Q, Chen S, Ge B.Detecting Salmonella Serovars in shell eggs by loop-mediated isothermal amplification[J]. J Food Protect, 2013, 76(10):1790-1796.
[26] Isaia S, Panagiotis K.Evaluationof loop-mediated isothermal amplification fordetection of Toxoplasmagondii in water samples and comparative findings by polymerase chain reaction and immunofluorescence test(IFT)[J]. Diagn Micr Infec Dis, 2008, 62:357-365.
[27] Hiroka A, Moemi S, Hiroshi I, et al.Rapid identification of plasmodium-carrying mosquitoes using loop-mediated isothermal amplification[J]. Biochem Biophys Res Commun, 2008, 37:671-676.
[28] Wang LX, He L, Fang R, ey al. Loop-mediated isothermal amplification(LAMP) assay for detection of Theileria sergenti infection targeting the p33 gene[J]. Vet Parasitol, 2010, 17l:159-162.
[29] Matovu E, Kuepfer I, Boobo A, et al.Comparative detection of trypanosomal DNA by loop-mediated isothermal amplification and PCR from flinders technology associates cards spotted with patient blood[J]. J Clin Microbiol, 2010, 48(6):2087-2090.
[30] Nkouawa A, Sako Y, Li T, et al.A loop-mediated isothermal amplification method for a differential identification of Taenia tapeworms from human: application to a field survey[J]. Parasitol Int, 2012, 61: 723-725.
[31] Sako Y, Nkouawa A, Yanagida T, et al.Loop-mediated isothermal amplification method for a differential identification of human taenia tapeworms[J]. Methods Mol Biol, 2013,1039:109-120.
[32] Ludwig N, Rudi Fv.Detection of Fusarium graminearum DNA using a loop-mediated isothermal amplification(LAMP) assay[J]. Int J Food Microbiol, 2010, 140:183-191.
[33] Saito R, Misawa Y, Moriya K, et al.Development and evaluation of a loop-mediated isothermal amplification assay for rapid detection of Mycoplasma pneumoniae[J]. Int J Med Microbiol, 2005, 54(11):1037-1041.
[34] Li J, Minion FC, Petersen AC, et al.Loop-mediated isothermal amplification for rapid and convenient detection of Mycoplasma hyopneumoniae[J]. World J Microb Biotechnol, 2013, 29(4):607-616.
[35] Tao C, Zhang Q, Zhai S, et al.Detection of HbsAg and hATIII genetically modified goats (Caprahircus) by loop-mediated isothermal amplification[J]. Mol Biol Rep, 2013, 40(11): 6177-6182.