摘要
为研究禽流感病毒(AIV)DNA疫苗重组质粒在组织中的分布情况,本研究以AIV DNA疫苗pCAGGoptiHA 5HA基因为检测对象,分别建立检测AIV DNA疫苗重组质粒的SYBR Green Ⅰ实时定量PCR方法和TaqMan MGB实时定量PCR方法。经优化比较两种方法的特异性、敏感性、重复性和污染率。结果表明,两种方法的标准曲线线性关系均较好,相关系数均达到0.999;最低检测量为42 copies,特异性强;探针法的重复性优于染料法,污染率低于染料法,因此采用TaqMan MGB实时定量PCR方法检测AIV DNA疫苗重组质粒组织分布情况。
To detect the DNA vaccine of pCAGGoptiHA5 in immunized chicks,we established the SYBR Green Ⅰ quantitative real-time PCR（QPCR） and TaqMan MGB QPCR methods with the primers and TaqMan MGB probes designed according to the sequence of pCAGGoptiHA5.After compared specificity,sensibility,reproducibility and contamination,we found that the correlation was above 0.999 and the detection sensibility was 42 copies,but the reproducibility and contamination of TaqMan MGB QPCR were better than SYBR Green I QPCR.Therefore,TaqMan MGB QPCR was a better method to study the distribution and persistence of plasmid DNA vaccine pCAGGoptiHA5 in chickens.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2011年第3期203-207,共5页
Chinese Journal of Preventive Veterinary Medicine
基金
十一五国家科技支撑计划(2006BAD06A05)
国家863计划(2006AA10A205)
