期刊文献+

皮毛中5种病毒基因芯片检测方法的研究 被引量:2

Development of a Gene Chip Method to Detect Five Viruses in Fur
下载PDF
导出
摘要 根据GenBank上蓝舌病病毒(BTV)、O型口蹄疫病毒(FMDV)、山羊痘病毒(GPV)、绵羊痘病毒(SPPV)和牛病毒性腹泻病毒(BVDV)等5种病毒的特异性保守序列,分别设计多重荧光标记引物和相应寡核苷酸探针。使用芯片点样液稀释各探针至终浓度30μmol/L,点样制备11×11阵列芯片。核酸杂交后,建立并优化基因芯片检测方法。结果显示,使用470mL/L甲酰胺杂交液,42℃摇转杂交4h为最佳基因芯片杂交条件。建立的基因芯片检测方法与伪狂犬病病毒(PRV)、猪繁殖与呼吸综合征病毒(PRRSV)、禽流感病毒(AIV)和新城疫病毒(NDV)等无交叉反应,检测敏感性可达20拷贝病毒核酸。制备的基因芯片稳定,保存6个月可用。对151份临床样品进行基因芯片和商业化PCR试剂盒平行检测,两者的符合率为100%。 Using the specific conservative sequences of BTV, FMDV/O,GPV,SPPV and BVDV which were searched in GenBank, we designed multiple PCR primers and corresponding specific oligo probes respectively. The probes were diluted to 30 μmol/L with dilution buffer and spotted on substrate-coated chip to 11 X 11 array. Followed by hybridization, the gene chip method was developed and optimized. The results showed that the effiident hybridization conditions were rotated 2 hours in 470 mL/L formamide at 42℃. The gene chip method had no cross-reactivity with PRV, PRRS, AIV and NDV. The gene chip stored for six months had a stable result and the sensitivity was up to 20 copies. The gene chip method and commer- cial PCR kit were used to detect the 151 clinical samples parallelly, and the coincidence was 100%.
出处 《动物医学进展》 CSCD 北大核心 2013年第4期27-32,共6页 Progress In Veterinary Medicine
基金 中国检科院基本科研业务费专项资金资助项目(2009JK010)
关键词 皮毛 动物病毒 基因芯片 检测 animal fur animal viruses gene chip detection
  • 相关文献

参考文献12

  • 1Bora D P, Venkatesan G, Bhanuprakash V, et al. TaqMan re- M-time PCR assay based on DNA polymerase gene for rapid detection of Off infection[J]. J Virol Meth, 2011, 178(1-2):249-252.
  • 2Batten C A, Bachanek-Bankowska K, Bin-Tarif A, et al. Bluetongue virus: European Community interqaboratory com- parison tests to evaluate ELISA and RT PCR detection meth ods[J]. Vet Mierobiol, 2008, 129(1-2) :80:88.
  • 3Stenfeldt C, Belsham G J. Detection of foot-and-mouth disease virus RNA in pharyngeal epithelium biopsy samples obtained from infected cattle: Investigation of possible sites of virus replication and persistence[J]. Vet Microbiol, 2012, 154 (3 4) :230-239.
  • 4Dubovi E J. Laboratory diagnosis of bovine viral diarrhea virus [J]. Biologicals, 2012, 6(4) :1-6.
  • 5Southern E, Mir K, Shchepinov M. Molecular interactions on microarrays[J]. Nat Gene, 1999, 21(1) : 5-9.
  • 6llatushna V J, Weller, Gibas C. Secondary structure in the target as a confounding factor in synthetic oligomer microarray design[J]. BMC Genomics, 2005, 6(1) :31.
  • 7Tian J, Ma K, Saaem I. Advancing high-throughput gene syn- thesis technology[J]. Molecular Bio Systems, 2009, 7 (5) .- 714-722.
  • 8Guo Z, Guilfoyle R A, Thiel A J, et al. Direct fluorescence a nalysis of genetic polymorphisms by hybridization with oligo nucleotide arrays on glass supports[J]. Nucleic Acids Res, 1994, 22(24) :5456-5465.
  • 9Rodaree K, Maturos T, Chaotheing S, et al. DNA hybridiza tion enhancement using piezoelectric microagitation through a liquid coupling medium[J]. Lab on a Chip, 2011,6(11): 1059-1064.
  • 10Brown P O, Botstein D. Exploring the new world of the ge nome with DNA microarrays[J]. Nat Gene, 1999, 21(1) : 33-37.

同被引文献19

引证文献2

二级引证文献3

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