摘要
目的 探讨巢式PCR技术检测人微小病毒B19(B19病毒 )非结构蛋白DNA的灵敏性及特异性。方法 参照经典的巢式PCR技术检测B19病毒结构蛋白DNA的方法 ,检测 30例孕妇血清及胎盘绒毛组织中的B19病毒结构蛋白及非结构蛋白DNA。并与人巨细胞病毒、乙肝病毒、丙肝病毒、腺病毒等及人类基因组DNA同批作巢式PCR检测 ,以观察巢式PCR检测B19病毒非结构蛋白DNA的特异性。结果 (1)巢式PCR检测B19病毒非结构蛋白DNA的最小检出量为 0 0 0 5fg ,灵敏性可达 0 0 0 5fg。 (2 )B19病毒质粒模板在 10 3bp处出现靶条带 ,而人巨细胞病毒、乙肝病毒、丙肝病毒、腺病毒等及人类基因组DNA均无扩增产物产生。 (3)孕妇血清及胎盘、绒毛组织中的B19病毒结构蛋白检出率均为 2 6 7% (8/30 ) ,而非结构蛋白DNA检出率在血清中为 33 3% (10 /30 ) ,胎盘、绒毛组织中均为 2 6 7% (8/30 )。两者比较 ,差异均无显著性 (P >0 0 5 )。结论 巢式PCR技术检测B19病毒非结构蛋白DNA是一种灵敏、特异、简便、快速诊断B19病毒感染的新方法。
Objective To explore the sensitivity and specificity of the new nested-polymerase chain reaction(nested-PCR) assay in the detection of human parvovirus B19(HPV B19) nonstructural protein DNA and the applicability of this technique in detecting the gravida infected with parvovirus B19. Methods Evaluating the new nested-polymerase chain reaction assay by routine process, and compared with the general nested-PCR in detecting structural protein DNA.Then, the two techniques were used to detect human parvovirus B19 structural protein and nonstructural protein DNA in gravida serum and pregnant tissue of 30 cases. Results The sensitivity of the new method was 0.005 fg which was similar to the general nested- polymerase chain reaction assay, and the detection results of other viruses and human genome DNA were all negative. The positive rates of structural protein DNA and nonstructural protein DNA in gravida sera were 26.7% and 33.3%, respectively. And the difference was non significant in statistics. At the same time, both of these two positive rates in placenta and villus were 26.7%. Conclusions The new nested-polymerase chain reaction assay was a sensitive, specific and convenient method in detecting human parvovirus B19 infection, and the detection of nonstructural protein DNA paves the way for the following study on the disease mechanisms of B19.
出处
《中华妇产科杂志》
CAS
CSCD
北大核心
2004年第2期86-90,共5页
Chinese Journal of Obstetrics and Gynecology
