目的建立快速、特异性好、灵敏度高的Real-Time PCR方法定量检测沙门菌。方法根据编码沙门菌肠毒素基因stn的核苷酸序列,设计荧光探针和一对引物,通过对荧光定量PCR反应体系和反应条件的摸索,建立定量检测沙门菌的方法。结果建立的Real-...目的建立快速、特异性好、灵敏度高的Real-Time PCR方法定量检测沙门菌。方法根据编码沙门菌肠毒素基因stn的核苷酸序列,设计荧光探针和一对引物,通过对荧光定量PCR反应体系和反应条件的摸索,建立定量检测沙门菌的方法。结果建立的Real-Ti me PCR方法有很好的特异性与敏感性,所检测沙门菌结果均为阳性,而非沙门菌均为阴性;标准曲线相关系数为R2=0.993,其敏感性为5CFU。运用该方法对108份鸡粪便、50份鸡肉以及58份水样进行检测,阳性率分别为3.7%(6/108)、4%(2/50)和3.4%(2/58),与传统细菌分离检测结果相符。结论结果表明该方法具有简便、快速、特异性强、敏感性高等特点,此研究为环境及疾病诊断中沙门菌快速检测提供了新方法。展开更多
By mating transgenic mice, transgenic mice model of high expression of foreign gene was screened. Making use of conventional way of breeding to mate 2(A♀,B♂)transgenic mice integrated Hepatitis B virus(HBV) gene, 2A...By mating transgenic mice, transgenic mice model of high expression of foreign gene was screened. Making use of conventional way of breeding to mate 2(A♀,B♂)transgenic mice integrated Hepatitis B virus(HBV) gene, 2A(♀♂) and 2B(♀♂)transgenic homozygote mice, and 6 bi-heterozygote mice(4♀2♂) by mating A and B homozygote mice were taken. Blood serum of these transgenic mice and their off-springs’ were collected, and HBV DNA expressing in blood serum of transgenic mice was tested by polymerase chain reaction and ELISA (PCR- ELISA). The results were as follows: HBV DNA expression in the blood of bi-heterozygote mice was much more than that of homozygote and heterozygote mice. Breeding bi-heterzygote mice is likely to be a good way of high expression of foreign gene in transgenic animals.展开更多
文摘目的建立快速、特异性好、灵敏度高的Real-Time PCR方法定量检测沙门菌。方法根据编码沙门菌肠毒素基因stn的核苷酸序列,设计荧光探针和一对引物,通过对荧光定量PCR反应体系和反应条件的摸索,建立定量检测沙门菌的方法。结果建立的Real-Ti me PCR方法有很好的特异性与敏感性,所检测沙门菌结果均为阳性,而非沙门菌均为阴性;标准曲线相关系数为R2=0.993,其敏感性为5CFU。运用该方法对108份鸡粪便、50份鸡肉以及58份水样进行检测,阳性率分别为3.7%(6/108)、4%(2/50)和3.4%(2/58),与传统细菌分离检测结果相符。结论结果表明该方法具有简便、快速、特异性强、敏感性高等特点,此研究为环境及疾病诊断中沙门菌快速检测提供了新方法。
文摘By mating transgenic mice, transgenic mice model of high expression of foreign gene was screened. Making use of conventional way of breeding to mate 2(A♀,B♂)transgenic mice integrated Hepatitis B virus(HBV) gene, 2A(♀♂) and 2B(♀♂)transgenic homozygote mice, and 6 bi-heterozygote mice(4♀2♂) by mating A and B homozygote mice were taken. Blood serum of these transgenic mice and their off-springs’ were collected, and HBV DNA expressing in blood serum of transgenic mice was tested by polymerase chain reaction and ELISA (PCR- ELISA). The results were as follows: HBV DNA expression in the blood of bi-heterozygote mice was much more than that of homozygote and heterozygote mice. Breeding bi-heterzygote mice is likely to be a good way of high expression of foreign gene in transgenic animals.
