风疹病毒宫内感染的实验诊断研究

目的：建立准确、快速的风疹病毒内感染实验诊断方法。方法：将疫苗株病毒稀释后接种Vero细胞，收集不同时段培养上清液和细胞，RT-PCR法检测风疹病毒RNA（培养RT-PCR法）。用直接RT—PCR法和培养RT—PCR检测可疑风疹病毒感染的胎儿组织和羊水标本。结果：直接RT—PCR法可检测模拟羊水标本中15TCID50／ml风疹病毒RNA。培养RT—PCR法在10TCID50的病毒感染后6h的V细胞和感染后24h的培养上清液中检出风疹病毒RNA。直接RT—PCR法和培养RT—PCR法检测4例足月产胎儿羊水均阴性；1例引产胎儿羊水直接RT—PCR法阴性，培养RT—PCR法在接种细胞后24h检出风疹病毒RNA。结论：培养RT—PCR法是一个快速、特异、灵敏的实验室诊断风疹病毒宫内感染的方法。

A METHODOLOGY RESEARCH ON PRENATAL DIAGNOSIS OF INTRAUTERINE RUBELLA VIRUS INFECTION

Objective:To develope a reliable and rapid method for prenatal diagnosis of intrauterine rubella virus infection.Methods:Vero cells were infected by rubella vaccine virus,and the supernatant and cells were collected at given time for RNA extraction and detected by RT-PCR(culture-RT-PCR).Tissues and amniotic fluid of high suspected fetus of intrauterine rubella virus infection were detected by direct RT-PCR and culture-RT-PCR.Results:The direct RT-PCR gained positive results when the titer of model samples was 15 TCID_(50)/ml and above.The culture-RT-PCR detected the virus RNA in 5 h' cells and 24 h' supernatant post infection of vero cells infected by 10 TCID_(50) of rubella vaccine virus.The culture-RT-PCR had the positive result at 6 h' Vero cells and 24 h' supernatant post infection of 10 TCID_(50) and above.Employing direct RT-PCR and cuture RT-PCR,4 term birth' amniotic fluid were all negative of rubella virus RNA.One induced amniotic fluid had a positive result by culture-PT-PCR at 24 h' cells post inoculation,while the direct RT-PCR was negative.Conclusion:Culture-RT-PCR is a rapid,specific and sensitive method to diagnose the intrauterine rubella virus infection.