实时荧光定量PCR对猪瘟疫苗中病毒含量的检测
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摘要
为了更准确地检测猪瘟疫苗中的病毒含量,为猪瘟免疫预防工作提供科学参考,试验设计针对CSFV E2基因的引物,利用RT-PCR方法扩增出目的片段,运用检测猪瘟病毒的SYBR Green-Ⅱ荧光定量PCR方法对来自不同生产厂家的6种猪瘟疫苗的病毒含量进行定量检测。结果:不同厂家生产的猪瘟兔化弱毒疫苗存在效价差异,6种疫苗中的病毒含量由高到低为A>D>F>C>E>B,分别为4.52×10~4、2.36×10~4、2.02×10~4、1.70×10~4、1.08×10~4、6.18×10~3 copies/μL。
In order to detect the viral content of swine fever vaccine more accurately,it provides scientific reference for the prevention of swine fever vaccine immunization. Experimental design for primers of CSFV E2 gene,amplification of target fragments by RT-PCR method. The viral contents of 6 swine fever vaccines from different manufacturers were quantitatively detected by SYBR Green-Ⅱ fluorescence quantitative PCR assay for detection of classical swine fever virus. Results,the difference of potency of swine fever virus vaccine produced by different factories in rabbits. The viral content in 6 vaccines from different manufacturers was A>D>F>C>E>B from high to low,respectively 4.52×10~4、2.36×10~4、2.02×10~4、1.70×10~4、1.08×10~4、6.18×10~3 copies/μL.
In order to detect the viral content of swine fever vaccine more accurately,it provides scientific reference for the prevention of swine fever vaccine immunization. Experimental design for primers of CSFV E2 gene,amplification of target fragments by RT-PCR method. The viral contents of 6 swine fever vaccines from different manufacturers were quantitatively detected by SYBR Green-Ⅱ fluorescence quantitative PCR assay for detection of classical swine fever virus. Results,the difference of potency of swine fever virus vaccine produced by different factories in rabbits. The viral content in 6 vaccines from different manufacturers was A>D>F>C>E>B from high to low,respectively 4.52×10~4、2.36×10~4、2.02×10~4、1.70×10~4、1.08×10~4、6.18×10~3 copies/μL.
引文
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[10]龚雪蕊,李阿茜,刘洋,等.寨卡病毒、登革病毒、基孔肯雅病毒三重荧光定量RT-PCR检测方法的建立及评价[J].病毒学报,2018,34(1):52-58.
[11]爨淑楠,张才,孙先利,等.犬巴贝斯虫实时荧光定量PCR检测方法的建立[J].黑龙江畜牧兽医,2017(21):168-171.
[12]高晓波,黄东风,姚学军,等.实时荧光PCR在H7N9禽流感病毒监测中的应用[J].当代畜牧,2017(30):44-46.
[13]白泉阳,傅光华,程龙飞,等.鸭源基因Ⅸ型禽1型副黏病毒实时荧光定量RT-PCR方法的建立与应用[J].中国兽医学报,2017,37(10):1868-1873.
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[15]陈锴,姚华伟,王长江,等.荧光定量PCR作为猪瘟兔化弱毒疫苗效价检验替代方法的研究与应用[J].中国农业科学,2013,46(1):162-169.
[16]葛叶,张兴娟,朱庆虎,等.荧光定量RT-PCR方法与兔体定型热试验对于检测猪瘟兔化弱毒疫苗的平行关系[J].中国预防兽医学报,2011,33(9):699-703.
[2]吴请民.兽医传染病学[M].北京:中国农业大学出版社,2002:223-254.
[3]仇华吉,童光志,沈荣显.猪瘟兔化弱毒疫苗-半个世纪的回顾[J].中国农业科学,2005,38(8):1675-1685.
[4]李春红,唐满华,陈瑞爱.猪瘟兔化弱毒疫苗效检替代方法研究进展[J].当代畜牧,2014(36):34-39.
[5]唐红慧,邵国青.猪瘟兔化弱毒苗抗原量测定方法的探索及不同免疫程序的差异性比较研究[J].扬州大学学报,2010,3(4):23-25.
[6]朱子健,唐志芬,杨金雨,等.猪瘟兔化弱毒疫苗与猪圆环病毒2型灭活疫苗同时接种的免疫效果评价[J].中国预防兽医学报,2017,39(2):148-151.
[7]王伟,邹月红,李雪松,等.猪瘟石门系强毒株SYBR Green-Ⅰ染料法实时荧光定量PCR方法的建立[J].中国畜牧兽医,2011,38(3):104-107.
[8]Nellemann C,Vingg aard A M,Dalgaar d M,et al.Quantificat ion of antiandrog en effect determined by light cycler technology[J].Toxico Logy,2001(163):329-381.
[9]吴玲玲,李艳芬,闫江舟,等.荧光PCR直接检测食物中毒样品中A(B)型肉毒梭菌[J].中国卫生检验杂志,2018(2):163-165.
[10]龚雪蕊,李阿茜,刘洋,等.寨卡病毒、登革病毒、基孔肯雅病毒三重荧光定量RT-PCR检测方法的建立及评价[J].病毒学报,2018,34(1):52-58.
[11]爨淑楠,张才,孙先利,等.犬巴贝斯虫实时荧光定量PCR检测方法的建立[J].黑龙江畜牧兽医,2017(21):168-171.
[12]高晓波,黄东风,姚学军,等.实时荧光PCR在H7N9禽流感病毒监测中的应用[J].当代畜牧,2017(30):44-46.
[13]白泉阳,傅光华,程龙飞,等.鸭源基因Ⅸ型禽1型副黏病毒实时荧光定量RT-PCR方法的建立与应用[J].中国兽医学报,2017,37(10):1868-1873.
[14]袁亚男,刘文忠.实时荧光定量PCR技术的类型、特点与应用[J].中国畜牧兽医,2008,35(3):27-30.
[15]陈锴,姚华伟,王长江,等.荧光定量PCR作为猪瘟兔化弱毒疫苗效价检验替代方法的研究与应用[J].中国农业科学,2013,46(1):162-169.
[16]葛叶,张兴娟,朱庆虎,等.荧光定量RT-PCR方法与兔体定型热试验对于检测猪瘟兔化弱毒疫苗的平行关系[J].中国预防兽医学报,2011,33(9):699-703.