人博卡病毒荧光定量PCR方法(TaqMan探针法)的建立及其在血液制品污染情况调查中的应用

Development of PCR Method Using TaqMan Probe for the Detection of Human Bocavirus and Investigation of Pollution of Blood Products and Raw Plasma

关键词:  博卡病毒  血液制品  检测方法  人凝血因子Ⅷ

Key words:  human bocavirus(HBoV)  blood products  test method  human clotting factor Ⅷ

摘要:

目的 开发快捷方便的检测人博卡病毒（human bocavirus，HBoV）方法，并调查华南地区血液制品和原料血浆中的污染情况。方法 建立HBoV的TaqMan探针实时荧光定量PCR方法，并对相关血液制品企业提供的单人份血浆和原料血浆样品、人血白蛋白、静脉注射免疫球蛋白（intravenous immunoglobulin，IVIG）、人纤维蛋白原、人凝血因子Ⅷ及人纤维蛋白胶-人纤维蛋白原中HBoV的污染情况进行检测。结果 HBoV-DNA在原料血浆和人凝血因子Ⅷ样本中的检测率分别为5.2%和5.0%，但在其他血液制品中则没有检出。结论 本研究建立的基于TaqMan探针HBoV的实时荧光定量PCR方法可以很好地检测出原料血浆及血液制品中的HBoV，对于血液制品的病毒安全有重要意义。

Abstract:

OBJECTIVE To develop a quick and convenient method for detection of human bocavirus (HBoV), and investigate the pollution situation of blood products and raw plasma in south of China. METHODS Real time fluorescence quantitative PCR using TaqMan probe was established. Blood products collected from different manufacturer including original single plasma and raw plasma samples, serum albumin, intravenous injection of immune globulin protein (IVIG), fibrin raw and human clotting factor Ⅷ and human fibrinogen fibrin glue to human fibrin were checked. RESULTS The detection rate of HBoV-DNA in raw plasma samples and human clotting factor Ⅷ were 5.2% and 5.0% respectively. But no HBoV-DNA was detected in other blood products. CONCLUSION PCR method developed in this thesis is qualified to test HBoV-DNA in blood products and raw plasma, which is of great significance to the standardized production and medication safety.