H7亚型和N9亚型禽流感病毒RT-LAMP可视化检测方法的建立

doi:10.11843/j.issn.0366-6964.2015.07.014

Abstract

Abstract:

A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for rapid visual detection of H7 and N9 subtype avian influenza viruses (AIV).According to the conservative sequences of hemagglutinin (HA) and neuraminidase (NA) genes of H7 and N9 subtype AIV in GenBank，two sets of primers were designed，then the reaction conditions were optimized.The specificity and sensitivity of the RT-LAMP assay were evaluated.RT-LAMP amplification was completed in 1 h by incubating all of the reagents in a single tube by real-time turbidimeter at 63 ℃.The specific test demonstrated that the RT-LAMP assay was specific for detecting H7 and N9 subtypes of AIV，and no cross reaction with other subtypes of AIV and avian respiratory pathogens.The detection limit of the RT-LAMP assay was 10 copies using vitro transcription RNA of target genes.The newly developed RT-LAMP assay is specific，sensitive，simple，rapid and can identify H7，N9 and H7N9 subtype AIV visually，and it will be a very useful screening assay for prevention and control these diseases.

CLC Number:

S852.659.5

LUO Si-si，XIE Zhi-xun，XIE Li-ji，DENG Xian-wen，XIE Zhi-qin，HUANG Jiao-ling，ZENG Ting-ting. Visual Detection of H7 and N9 Subtypes Avian Influenza Viruses by Reverse Transcription Loop-mediated Isothermal Amplification Assay[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(7): 1176-1183.

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https://www.xmsyxb.com/EN/Y2015/V46/I7/1176

References

［1］CHAN J F，TO K K，TSE H，et al.Interspecies transmission and emergence of novel viruses：lessons from bats and birds［J］. Trends Microbiol，2013，21(10)：544-555.
［2］TONG S，ZHU X，LI Y，et al.New world bats harbor diverse influenza A viruses［J］.PLoS Pathog，2013，9 (10)：e1003657.
［3］BEIGEL J H，FARRAR J，HAN A M，et al.Avian influenza A (H5N1) infection in humans［J］. N Engl J Med，2005，353 (13)：1374-1385.
［4］PAPPAS C，MATSUOKA Y，SWAYNE D E，et al.Development and evaluation of an influenza virus subtype H7N2 vaccine candidate for pandemic preparedness［J］.Clin Vaccine Immunol，2007，14(11)：1425-1432.
［5］TWEED S A，SKOWRONSKI D M，DAVID S T，et al.Human illness from avian influenza H7N3，British Columbia［J］.Emerg Infect Dis，2004，10(12)：2196-2199.
［6］FOUCHIER R A，SCHNEEBERGER P M，ROZENDAAL F W，et al.Avian influenza A virus (H7N7) associated with human conjunctivitis and a fatal case of acute respiratory distress syndrome［J］.Proc Natl Acad Sci USA，2004，101(5)：1356-1361.
［7］PEIRIS M，YUEN K Y，LEUNG C W，et al.Human infection with influenza H9N2［J］.Lancet，1999，354(9182)：916-917.
［8］谭伟，谢芝勋.H10N8亚型禽流感病毒的研究进展［J］.中国畜牧兽医，2014，41(8)：236-241.
TAN W，XIE Z X.Research progress on avian influenza a virus subtype H10N8［J］.China Animal Husbandry & Veterinary Medicine，2014，41(8)：236-241.(in Chinese)
［9］QI X，CUI L，YU H，et al.Whole-Genome sequence of a reassortant H5N6 avian influenza virus isolated from a live poultry market in China，2013［J］.Genome Announc，2014，2(5).pii：e00706-00714.
［10］NOTOMI T，OKAYAMA H，MASUBUCHI H，et al.Loop-mediated isothermal amplification of DNA［J］. Nucleic Acids Res，2000，28(12)：E63.
［11］PENG Y，XIE Z X，LIU J，et al.Visual detection of H3 subtype avian influenza viruses by reverse transcription loop-mediated isothermal amplification assay［J］.Virol J，2011，8：337.
［12］XIE Z，TANG Y，FAN Q，et al.Rapid detection of group I avian adenoviruses by a loop-mediated isothermal amplification［J］.Avian Dis，2011，55(4)：575-579.
［13］XIE Z，FAN Q，LIU J，et al.Reverse transcription loop-mediated isothermal amplification assay for rapid detection of Bovine Rotavirus［J］.BMC Vet Res，2012，8：133.
［14］罗思思，谢芝勋，邓显文，等.鸡毒支原体强弱毒株LAMP可视化鉴别检测方法的建立［J］.中国兽医科学，2012，42（9）：943-948.
LUO S S，XIE Z X，DENG X W，et al.Development of a loop-mediated isothermal amplification assay for visual differentiation of Mycoplasma gallisepticum virulent strains from attenuated strains［J］.Chinese Veterinary Science，2012，42（9）：943-948.(in Chinese)
［15］罗思思，谢芝勋，刘加波，等.H7N9亚型禽流感病毒三重PCR检测方法的建立［J］.中国畜牧兽医，2014，41（2）：61-64.
LUO S S，XIE Z X，LIU J B，et al.Development of a triplex PCR assay for detection of H7N9 subtype avian influenza virus［J］.China Animal Husbandry & Veterinary Medicine，2014，41（2）：61-64.(in Chinese)
［16］罗宝正，莫秋华，李儒曙，等.新型H7N9亚型禽流感病毒多重荧光RT-PCR快速检测方法的建立［J］.病毒学报，2014，30（1）：1-5.
LUO B Z，MO Q H，LI R S，et al.Rapid detection of novel avian influenza virus subtype H7N9 by multiplex real-time RT-PCR［J］.Chinese Journal of Virology，2014，30（1）：1-5.(in Chinese)
［17］刘业兵，张磊，孙跃辉，等.可视化的RT-LAMP方法检测禽白血病病毒［J］.畜牧兽医学报，2011，42 (1)：150-156.
LIU Y B，ZHANG L，SUN Y H，et al.Visualized detection of avian leukosis virus by reverse transcription loop-mediated isothermal amplification assay［J］.Acta Veterinaria et Zootechnica Sinica，2011，42(1)：150-156.(in Chinese)
［18］李兆龙，陈仕龙，林锋强，等.禽新型黄病毒RT-LAMP检测方法的建立［J］.畜牧兽医学报，2012，43（4）：659-663.
LI Z L，CHEN S L，LIN F Q，et al.Establishment of rapid reverse transcription loop mediated isothermal amplification for detection of avian novel flavivirus［J］.Acta Veterinaria et Zootechnica Sinica，2012，43（4）：659-663.(in Chinese)
［19］蒋菲，黄书林，徐威，等.H9亚型禽流感病毒环介导等温扩增检测方法的建立与荧光指示剂的应用［J］.农业生物技术学报，2011，19(1)：191-196.
JIANG F，HUANG S L，XU W，et al.Detection of H9 avian influenza virus by loop-mediated isothermal amplification and application of fluorescent reagent［J］.Journal of Agricultural Biotechnology，2011，19(1)：191-196.(in Chinese)
［20］罗剑鸣，吴希阳，徐子乾，等.基于颜色判定的逆转录环介导等温扩增技术检测GII型诺如病毒基因［J］.病毒学报，2012，28(2)：165-171.
LUO J M，WU X Y，XU Z Q，et al.Colorimetric detection of norovirus genotype GII by reverse transcription loop-mediated isothermal amplification［J］.Chinese Journal of Virology，2012，28(2)：165-171.(in Chinese)
［21］XIE L，XIE Z，ZHAO G，et al.A loop-mediated isothermal amplification assay for the visual detection of duck circovirus［J］.Virol J，2014，11：76.
［22］薛超波，黄朱梁，孙瑛，等.海产品中创伤弧菌实时浊度LAMP检测方法的建立［J］.中国预防兽医学报，2013，35(11)：912-915.
XUE C B，HUANG Z L，SUN Y，et al.Development of a real-time turbidimeter-based loop-mediated isothermal amplification for detection of Vibrio vulnificus in seafood［J］.Chinese Journal of Preventive Veterinary Medicine，2013，35(11)：912-915.(in Chinese)
［23］袁瑛娜，单潇潇，王宗德，等.应用LAMP实时浊度法检测转基因大豆［J］.现代食品科技，2011，27(10)：1264-1267.
YUAN Y N，SHAN X X，WANG Z D，et al.Development of a real-time turbidimeter-based loop-mediated isothermal amplification assay for detection of transgenic soybean［J］.Modern Food Science and Technology，2011，27(10)：1264-1267.(in Chinese)
［24］董志珍，郑文杰，王乃福，等.H7N9禽流感病毒等温扩增快速检测方法的建立［J］.中国动物检疫，2013，30（7）：72-77.
DONG Z Z，ZHENG W J，WANG N F，et al.Development of rapid assay for H7N9 influenza virus by loop-mediated isothermal amplification method［J］.China Animal Health Inspection，2013，30（7）：72-77.(in Chinese)

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Visual Detection of H7 and N9 Subtypes Avian Influenza Viruses by Reverse Transcription Loop-mediated Isothermal Amplification Assay

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