• Chinese Core Journal
  • Chinese Science Citation Database (CSCD) Source journal
  • Journal of Citation Report of Chinese S&T Journals (Core Edition)
Advanced Search
JIA Kun, HAN Taiguang, YUAN Liguo, SUN Lingshuang, NING Zhangyong, WANG Heng, LI Shoujun. Establishment and application of visual RT-LAMP technology for detecting bovine ephemeral fever virus[J]. Journal of South China Agricultural University, 2017, 38(3): 15-20. DOI: 10.7671/j.issn.1001-411X.2017.03.003
Citation: JIA Kun, HAN Taiguang, YUAN Liguo, SUN Lingshuang, NING Zhangyong, WANG Heng, LI Shoujun. Establishment and application of visual RT-LAMP technology for detecting bovine ephemeral fever virus[J]. Journal of South China Agricultural University, 2017, 38(3): 15-20. DOI: 10.7671/j.issn.1001-411X.2017.03.003
shu

Establishment and application of visual RT-LAMP technology for detecting bovine ephemeral fever virus

  • College of Veterinary Medicine, South China Agricultural University/Key Laboratory of Prevention and Control for Severe Clinical Animal Diseases of Guangdong Province/Guangdong Technological Engineering Research Center for Pet, Guangzhou 510642, China

More Information
  • Received Date: June 20, 2016
  • Available Online: May 17, 2023
    Graphical Abstract
    • Abstract
  • Objective 

    To develop a rapid, convenient and visual molecular assay for detecting bovine ephemeral fever virus (BEFV).

    Method 

    Two pairs of primers based on six conserved regions in G protein gene of BEFV were designed, and the visual reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for detecting BEFV was developed. The reaction conditions of RT-LAMP assay were optimized, and the assay was compared with PCR.

    Result 

    The reaction ladder bands from the RT-LAMP assay were most obvious when the RT-LAMP reaction system contained 3 mmol·L-1 Mg2+, 0.4 mol·L-1 betaine and 1.2 μmol·L-1 dNTPs mix, the ratio of inner to outer primers was 8:1, and the reaction temperature was 63 ℃. Clear reaction ladder bands were observed after 40 min of reaction. The established RT-LAMP assay had excellent specificity with only BEFV being amplified. The sensitivity was 10 times higher than that of ordinary PCR.

    Conclusion 

    The visual RT-LAMP assay is easy to operate and highly specific, and the results can be conveniently determined. This method can be used for the rapid detection of bovine epidemic fever.

    • FullText(HTML)
    • References (15)
  • [1]
    NANDI S, NEGI B S. Bovine ephemeral fever: A review[J]. Comp Immunol Microbiol Infect Dis, 1999, 22(2): 81-91. doi: 10.1016/S0147-9571(98)00027-7
    [2]
    AZIZ-BOARON O, KLAUSNER Z, HASOKSUZ M, et al. Circulation of bovine ephemeral fever in the Middle East--strong evidence for transmission by winds and animal transport[J]. Vet Microbiol, 2012, 158(3/4): 300-307.
    [3]
    DAVIES F G, OCHIENG P, WALKER A R. The occurrence of ephemeral fever in Kenya, 1968-1988[J]. Vet Microbiol, 1990, 22(2/3): 129-136.
    [4]
    ZHENG F, LIN G, ZHOU J, et al. A reverse-transcription, loop-mediated isothermal amplification assay for detection of bovine ephemeral fever virus in the blood of infected cattle[J]. J Virol Methods, 2011, 171(1): 306-309. doi: 10.1016/j.jviromet.2010.10.028
    [5]
    LI Z, ZHENG F, GAO S, et al. Large-scale serological survey of bovine ephemeral fever in China[J]. Vet Microbiol, 2015, 176(1/2): 155-160. https://www.deepdyve.com/lp/elsevier/large-scale-serological-survey-of-bovine-ephemeral-fever-in-china-vSyWodWWMs
    [6]
    吴清明, 魏文才, 陈水生, 等.奶牛流行热疫情的防控应对[J].南方奶业, 2013(1): 11-13.
    [7]
    白文彬, 姜春凌, 严隽端, 等.牛流行热病毒微量血清中和试验[J].中国畜禽传染病, 1988(4): 40-43. http://www.cnki.com.cn/Article/CJFDTOTAL-ZGXQ198804018.htm
    [8]
    ZHENG F Y, LIN G Z, QIU C Q. Development and application of G1-ELISA for detection of antibodies against bovine ephemeral fever virus[J]. Res Vet Sci, 2009, 87(2): 211-212. doi: 10.1016/j.rvsc.2009.03.010
    [9]
    STRAM Y, KUZNETZOVA L, LEVIN A. A real-time RT-quantative(q)PCR for the detection of bovine ephemeral fever virus[J]. J Virol Methods, 2005, 130(1/2): 1-6. doi: 10.1007/s12250-007-0031-2
    [10]
    BLASDELL K R, ADAMS M M, DAVIS S S. A reverse-transcription PCR method for detecting all known ephemeroviruses in clinical samples[J]. J Virol Methods, 2013, 191(2): 128-135. doi: 10.1016/j.jviromet.2013.04.011
    [11]
    刘志玲, 陈茹, 邱索平, 等.牛流行热病毒LUXTM荧光RT-PCR检测方法的建立[J].生物技术通报, 2010 (8): 174-179. http://www.cnki.com.cn/Article/CJFDTOTAL-SWJT201008037.htm
    [12]
    NOTOMI T, OKAYAMA H, MASUBUCHI H, et al. Loop-mediated isothermal amplification of DNA[J]. Nucleic Acids Res, 2000, 28(12): 63. doi: 10.1093/nar/28.12.e63
    [13]
    CHEN C, CUI S. Detection of porcine parvovirus by loop-mediated isothermal amplification[J]. J Virol Methods, 2009, 155(2): 122-125. doi: 10.1016/j.jviromet.2008.10.004
    [14]
    SHIVAKOTI S, ITO H, MURASE T, et al. Development of reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay for detection of avian influenza viruses in field specimens[J]. J Vet Med Sci, 2010, 72(4): 519-523. doi: 10.1292/jvms.09-0473
    [15]
    ZHU R Y, ZHANG K X, ZHAO M Q. Use of visual loop-mediated isotheral amplification of rimM sequence for rapid detection of Mycobacterium tuberculosis and Mycobacterium bovis [J]. J Microbiol Methods, 2009, 78(3): 339-343. doi: 10.1016/j.mimet.2009.07.006
    • Related Articles

  • Related Articles

    [1]CHU Zhizhan, WU Guanghong, LIU Xiaolin. A research on extraction and enzymatic hydrolysis of zein[J]. Journal of South China Agricultural University, 2014, 35(3): 86-89. DOI: 10.7671/j.issn.1001-411X.2014.03.016
    [2]BAI Wenjuan, HU Rongrong, ZHANG Jiaen, FENG Lifang, XU Huaqin. A Study on the Isolation, Screening and Phosphate Solubilizing Capacity of Phosphate Solubilizing Bacteria in the Rhizosphere of Corn[J]. Journal of South China Agricultural University, 2013, 34(2): 167-176. DOI: 10.7671/j.issn.1001-411X.2013.02.009
    [3]YIN Wei, LU Ying, GAN Hai-hua, CHEN Chun-xia, DONG Fei. Assessment of Chemical Remediation Effect on Pb-Contaminated Urban Soil by Using in vitro Test[J]. Journal of South China Agricultural University, 2011, 32(4): 27-30. DOI: 10.7671/j.issn.1001-411X.2011.04.007
    [4]CHEN Zhi-qiang,GUO Tao,LIU Yong-zhu,WANG Hui. Achievements and Perspective Outlook of Rice Space Induced Mutation Breeding in China[J]. Journal of South China Agricultural University, 2009, 30(1). DOI: 10.7671/j.issn.1001-411X.2009.01.002
    [5]ZHANG Yi-min,SUN Ya-kai,L Xue-bin,DAN Han-bin,WANG Hong. Isolation of the High Efficiency Phosphate Solubilization Bacterium Bmp5 and Its Activity and Culture Conditions[J]. Journal of South China Agricultural University, 2006, 27(3): 61-65. DOI: 10.7671/j.issn.1001-411X.2006.03.017
    [6]ZHANG Yi-ping,LIU Yang. A comparative research on microclimate characteristics between ancient tea plantation and conventional tea plantation in Yunnan Province[J]. Journal of South China Agricultural University, 2005, 26(2): 17-21. DOI: 10.7671/j.issn.1001-411X.2005.02.005
    [7]TANG Jin-chi~. Comparison of potassium efficiency of two soybean genotypes[J]. Journal of South China Agricultural University, 2005, 26(1): 7-10. DOI: 10.7671/j.issn.1001-411X.2005.01.002
    [8]QIAO Xi-ming,ZHANG Xian-feng,ZHU Zhong-shi. Experimental research on the components of minn-stripper harvester[J]. Journal of South China Agricultural University, 2004, 25(4): 98-101. DOI: 10.7671/j.issn.1001-411X.2004.04.025
    [9]Study on Influence of Controlled-Release Fertilizers to N Loss Leaching and Rice Yield[J]. Journal of South China Agricultural University, 2002, 23(3): 91-91. DOI: 10.7671/j.issn.1001-411X.2002.03.026
    [10]WU Ping-xiao,LIAO Zong-wen,Mao Xiao-yun. Research to the Fertilizer Efficiency of Modified Ammonium Bicarbonate and Its Eluviation Characteristics(Preliminary Report)[J]. Journal of South China Agricultural University, 2000, (3): 94-94. DOI: 10.7671/j.issn.1001-411X.2000.03.026

Catalog

    Get Citation
    PDF
    XML
    Article views (1437) PDF downloads (1827) Cited by()
    Turn off MathJax
    Article Contents
    Abstract
    References
    Related Articles

    Address： No. 483 Wushan Road, Tianhe District, Guangzhou Editorial Department of Journal of South China Agricultural University

    Postal Code：510642

    Tel：(020) 85280069/38746672

    Sponsor/Sponsored by：South China Agricultural University

    Editor-in-chief：Xue Hongwei

    ISSN: 1001-411X CN: 44-1110/S

    Email Alert
    RSS

    Supported by: Beijing Renhe Information Technology Co., Ltd.  Baidu Analytics

    /

    DownLoad:  Full-Size Img  PowerPoint
    Return
    Return