食源性致病菌等温扩增检测技术的研究进展
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摘要
快速可靠的食源性致病菌检测方法是保障食品安全的关键。然而,传统培养法的检测周期过于冗长,无法适应对时间敏感度较高的现场或在线检测。为此,研究人员开发了多种以快速和高灵敏度为主要特征的等温扩增技术及相应的检测产品,为食源性致病菌的现场或在线检测提供了强有力的技术支撑。该文对环介导等温扩增、滚环扩增、链置换扩增、切口酶信号扩增、核酸外切酶Ⅲ辅助扩增5种等温扩增方法的原理及其优缺点进行了梳理和比较。在此基础上,分析了等温扩增方法在细菌活性识别、非特异性扩增、前处理效率、检测通量4个方面所遇到的共性问题,并结合作者已有工作,提出了解决这些问题的方法或策略。
Rapid and reliable detection methods for food-borne pathogen are the key points for food safety control.However,traditional culture-based procedure for bacterial identification,generally taking 5 to 7 days and requiring skilled person and non-portable equipment,is incompetent for point-of-care or on-site detection.Thus,several isothermal amplification methods characterized by rapidness and thermal cycle independent were developed to fulfill some specific needs for point-of-care or on-site detection.In this review,mechanisms for isothermal amplification methods,designated as loop-mediated isothermal amplification,rolling circle amplification,strand displacement amplification,nicking enzyme signal amplification,exonuclease Ⅲ assisted amplification,were explained and compared in detail.Besides,some drawbacks shared by these methods were highlighted,and ways to overcoming those drawbacks such as adoption of propidium monoazide,novol fluorescent dye,high-throughput pretreatment equipment and microfluidic chip were provided based on the recent publications of the author?s group and others.
Rapid and reliable detection methods for food-borne pathogen are the key points for food safety control.However,traditional culture-based procedure for bacterial identification,generally taking 5 to 7 days and requiring skilled person and non-portable equipment,is incompetent for point-of-care or on-site detection.Thus,several isothermal amplification methods characterized by rapidness and thermal cycle independent were developed to fulfill some specific needs for point-of-care or on-site detection.In this review,mechanisms for isothermal amplification methods,designated as loop-mediated isothermal amplification,rolling circle amplification,strand displacement amplification,nicking enzyme signal amplification,exonuclease Ⅲ assisted amplification,were explained and compared in detail.Besides,some drawbacks shared by these methods were highlighted,and ways to overcoming those drawbacks such as adoption of propidium monoazide,novol fluorescent dye,high-throughput pretreatment equipment and microfluidic chip were provided based on the recent publications of the author?s group and others.
引文
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[2] National Health and Family Planning Commission of PRC.China Health and Family Planning Statistical Yearbook.Beijing:Peking Union Medical College Press(国家卫生和计划生育委员会.中国卫生和计划生育统计年鉴.北京:中国协和医科大学出版社),2017.
[3] Zhang S H,Guo L Z,Yang K,Zhang Y,Ye C S,Chen S,Yu X,Huang W E,Cui L.Front.Microbiol.,2018,9:2243.
[4] Li Q,Fang J H,Liu X,Xi X,Li M,Gong Y,Zhang M Z.Eur.Food Res.Technol.,2013,236(4):589-598.
[5] Ma B,Dai M Y,Fang J H,Wu Y Y,Zhang M Z.Am.J.Food Technol.,2016,11(5):193-203.
[6] Zhong J L,Zhao X H.Food Anal.Methods,2018,11(6):1543-1560.
[7] Notomi T,Okayama H,Masubuchi H,Yonekawa T,Watanabe K,Amino N,Hase T.Nucleic Acids Res.,2000,28(12):e63.
[8] Jaroenram W,Cecere P,Pompa P P.J.Microbiol.Methods,2018,doi:10.1016/j.mimet.2018.11.020.
[9] Wachiralurpan S,Sriyapai T,Areekit S,Kaewphinit T,Sriyapai P,Santiwatanakul S,Chansiri K.Food Anal.Methods,2017,10(11):3763-3772.
[10] Garrido-Maestu A,Azinheiro S,Carvalho J,Fucinos P,Prado M.Food Control,2018,86:27-34.
[11] Feng J L,Dai Z Y,Tian X L,Jiang X N.Food Control,2018,85:443-452.
[12] Kaur A,Kapil A,Elangovan R,Jha S,Kalyanasundaram D.PLoS One,2018,13:e0194817.
[13] Fang J H,Wu Y Y,Qu D,Ma B,Yu X P,Zhang M Z,Han J.Lett.Appl.Microbiol.,2018,67:79-88.
[14] Chun H J,Kim S,Han Y D,Kim K R,Kim J H,Yoon H,Yoon H C.ACS Sensors,2018,3:2261-2268.
[15] Sabike I I,Uemura R,Kirino Y,Mekata H,Sekiguchi S,Farid A S,Goto Y,Horii Y,Yamazaki W.Food Control,2017,74:107-111.
[16] Sabike I I,Uemura R,Kirino Y,Mekata H,Sekiguchi S,Okabayashi T,Goto Y,Yamazaki W.Front.Microbiol.,2016,7:1582.
[17] Romero M R,Cook N.Front.Microbiol.,2018,9:2401.
[18] Wang F,Jiang L,Ge B.J.Clin.Microbiol.,2012,50:91-97.
[19] Teng J,Ye Y,Yao L,Yan C,Cheng K,Xue F,Pan D,Li B,Chen W.Microchim.Acta,2017,184(9):3477-3485.
[20] Kong C,Wang Y,Fodjo E K,Yang G X,Han F,Shen X S.Microchim.Acta,2017,185:35.
[21] Chen S,Ge B.BMC Microbiol.,2010,10(1):1-9.
[22] Zhi A T,Ma B,Wu Y Y,Fang J H,Yu X P,Zhang M Z.Food Anal.Methods,2018,11:99-110.
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[37] Gerasimova Y V,Peck S,Kolpashchikov D M.Chem.Commun.,2010,46(46):8761-8763.
[38] Zhang Z Z,Zhang C Y.Anal.Chem.,2012,84(3):1623-1629.
[39] Sun Y,Zhao C,Yan Z,Ren J,Qu X.Chem.Commun.,2016,52:7505-7508.
[40] Zuo X,Xia F,Xiao Y,Plaxco K W.J.Am.Chem.Soc.,2010,132:1816-1818.
[41] Luo C,Tang H,Cheng W,Yan L,Zhang D,Ju H,Ding S.Biosens.Bioelectron.,2013,48:132-137.
[42] Wang W,Bao T,Zeng X,Xiong H,Wen W,Zhang X,Wang S.Biosens.Bioelectron.,2017,91:183-189.
[43] Chang Y C,Yang C Y,Sun R L,Cheng Y F,Kao W C,Yang P C.Sci.Rep.,2013,3:1863.
[44] Wang X R,Wu L F,Wang Y,Ma Y Y,Chen F H,Ou H L.Appl.Biochem.Biotechnol.,2015,175:882-891.
[45] Zhang P,Liu H,Ma S,Men S,Li Q,Yang X,Wang H,Zhang A.Biosens.Bioelectron.,2016,80:538-542.
[46] Zhang T T,Wang L,Gong P,Chen F X.Jiangsu Agric.Sci.(张涛涛,王兰,龚频,陈福欣.江苏农业科学),2014,42(2):238-241.
[47] Yamazaki W,Taguchi M,Kawai T,Kawatsu K,Sakata J,Inoue K,Misawa N.Appl.Environ.Microbiol.,2009,75(6):1597-1603.
[48] Lin W H,Zou B J,Song Q X,Zhou G H.Hereditas(林文慧,邹秉杰,宋沁馨,周国华.遗传),2015,37(9):899-910.
[49] Ma B,Wu Y Y,Dai M Y,Fang J H,Zhang M Z.Modern Food Sci.Technol.(马骉,吴莹莹,戴明雁,方结红,张明洲.现代食品科技),2016,32(7):205-213.
[50] Bosshard F,Berney M,Scheifele M,Weilenmann H U,Egli T.Microbiology,2009,155(4):1310-1317.
[51] Lin J,Ma B,Fang J H,Wang Y,He H,Lin W,Su W,Zhang M Z.Clin.Lab.,2017,63(3):495-505.
[52] Bakthavathsalam P,Longatte G,Jensen S O,Manefield M,Gooding J J.Sens.Actuators B,2018,268:255-263.
[53] Zhang M Z,Wang M Z,Chen Z L,Fang J H,Fang M M,Liu J,Yu X P.Anal.Bioanal.Chem.,2009,395(8):2591-2599.
[54] Kang M.Design and Implement for Fully Automatic Nucleic Acid Extraction System Based on Magnetic Separation.Nanjing:Southeast University(康淼.基于磁分离的全自动核酸提取系统设计与实现.南京:东南大学),2015.
[55] Zhou Q J,Wang L,Chen J,Wang R N,Shi Y H,Li C H,Zhang D M,Yan X J,Zhang Y J.J.Microbiol.Methods,2014,104:26-35.