期刊文献+

免疫磁珠富集与SYB RGreen荧光定量PCR技术联合检测水体中甲肝病毒 被引量:5

DETECTION ON HEPATITIS A VIRUS IN WATER SAMPLE USING IMMUNOMAGNETIC BEADS-BASED CONCENTRATION COMBINED WITH SYBR GREEN REAL-TIME PCR
原文传递
导出
摘要 本实验旨在建立一种免疫磁珠与SYBR Green荧光定量PCR技术联合检测水体中甲肝病毒的方法。通过制备特异性甲肝病毒免疫磁珠,优化病毒核酸提取方法,建立了免疫磁珠富集病毒和荧光定量PCR检测水体中甲肝病毒的方法。研究结果显示,1mg免疫磁珠与75μg甲肝病毒单克隆抗体偶联时,有最大抗体偶联量;该偶联后的磁珠富集甲肝病毒效率为92.87%。研究结果还显示,SYBR Green荧光定量PCR检测的标准曲线线性关系良好(R^2=0.999),检测极限低至9.27×10~1拷贝/μL。本研究结果表明,免疫磁珠与SYBR Green荧光定量PCR技术结合,具有特异性强、灵敏度高、可信度强、简便快捷的优点,本方法对水体中甲肝病毒污染状况检测与评价提供了有力的技术支持。 The aim of the experiment was to build up a detection method of hepatitis A virus in water sample,by using immunomagnetic beads- based and SYBR Green real- time PCR(IMB- real- time PCR).By preparing the magnetic beads coated with HAV monoclonal antibodies and optimizing the viral RNA extraction methods,the detection methods of hepatitis A virus in water sample were set up,such as utilizing immunomagnetic beads- based virus concentration and real- time PCR.The data showed that 1mg immunomagnetic beads which coated with 75μg HAV antibodies had the greatest efficiency(92.87%) to capture HAV in water sample.Meanwhile,the standard curve of real- time PCR had good linear relationship(reliability was 0.999).The limit of detection was improved as low as 9.27×10^1 copies/μL.The results indicated that the immunomagnetic beads- based with SYBR Green real- time PCR is a rapid,sensitive,specific and reliable method,which provides a strong technical basis for detection of HAV and evaluation in water.
出处 《内蒙古农业大学学报(自然科学版)》 CAS 北大核心 2014年第2期13-18,共6页 Journal of Inner Mongolia Agricultural University(Natural Science Edition)
基金 国家科技支撑计划(2013BAK12B05)
关键词 免疫磁珠 荧光定量PCR 甲肝病毒 Immunomagnetic beads Real-time PCR Hepatitis A virus
  • 相关文献

参考文献16

  • 1Martin A, Lemon S M. Hepatitis A virus: from discovery to vaccines [ J ]. Hepatology, 2006, 43 ( SI ) : S164 - $172. 被引量:1
  • 2杨万,何苗,李丹,施汉昌,刘丽.免疫磁珠分离与实时定量PCR技术联合检测水中轮状病毒的研究[J].环境科学,2009,30(5):1368-1375. 被引量:10
  • 3Yang W, Gu A Z, Zeng S, et al. Development of a com- bined immunomagnetic separation and quantitative reverse transcription - PCR assay for sensitive detection of infec- tious rotavirus in water samples [ J ]. Journal of microbio- logical methods, 2011, 84(3): 447-453. 被引量:1
  • 4徐金亭,李志清,向军俭,唐勇,杨红宇.单增李斯特菌免疫磁珠的制备研究[J].食品工业科技,2012,33(5):323-327. 被引量:15
  • 5Brooks H A, Gersberg R M, Dhar A K. Detection and quantification of hepatitis A virus in seawater via real - time RT - PCR[J].Journal of virological methods, 2005, 127(2): 109-118. 被引量:1
  • 6Villar L M, de Paula V S, Diniz - Mendes L, et al. Evalu- ation of methods used to concentrate and detect hepatitis Avirus in water samples[ J ]. Journal of virological methods, 2006, 137(2): 169-176. 被引量:1
  • 7Haramoto E, Katayama H, Oguma K, et al. Quantitative analysis of human enteric adenoviruses in aquatic environ- ments[J]. Journal of applied microbiology, 2007, 103 (6) : 2153 -2159. 被引量:1
  • 8Polaczyk A L, Narayanan J, Cromeans T L, et al. Uhrafil- tration - based techniques for rapid and simultaneous con- centration of multiple microbe classes from 100 - L tap wa- ter samples [ J ]. Journal of microbiological methods, 2008, 73 (2) : 92 - 99. 被引量:1
  • 9Bennett A R, MaePhee S, Betts R P. The isolation and detection of Escherichia coli O157 by use of immunomag- netic separation and immunoassay procedures[ J]. Leuers in applied microbiology, 1996, 22 (3) : 237 - 243. 被引量:1
  • 10Fratamico P M, Schuhz F J, Buchanan R L. Rapid iso- lation of Escherichia coli O157 : H7 from enrichment cul- turcs of foods using an immunomagnetic separation meth- od[J]. Food microbiology, 1992, 9(2): 105 -113. 被引量:1

二级参考文献48

共引文献33

同被引文献42

引证文献5

二级引证文献11

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部