Bosma T J, Corbett K M, Eckstein M B, O'Shea S, Vijayalakshmi P, Banatvala J E, Morton K, Best J M
Department of Virology, United Medical School, Guys Hospital, London, United Kingdom.
J Clin Microbiol. 1995 Nov;33(11):2881-7. doi: 10.1128/jcm.33.11.2881-2887.1995.
A reverse transcription-nested PCR assay (RT-PCR) was evaluated for diagnosis of congenitally acquired rubella in utero and during infancy. RT-PCR was compared with virus isolation for retrospective detection of rubella virus in placental and fetal tissues obtained after termination of pregnancy following primary rubella or rubella virus reinfection. Concordant results were obtained for 85% of samples; rubella virus RNA was detected by RT-PCR alone in four samples, and rubella virus was detected by isolation alone in two samples. Samples were also obtained for prenatal diagnosis of congenital infection; rubella virus RNA was detected in three of seven chorionic villus samples and one of three amniotic fluid samples by RT-PCR, while rubella virus was isolated in only one chorionic villus sample. To demonstrate that the RNA extracted from chorionic villus samples contained amplifiable RNA, a nested RT-PCR was used to detect keratin mRNA. Rubella virus was detected in placenta in two cases in which the fetus was uninfected, and there was no evidence of rubella virus in the placenta from one case in which the fetus was infected. Thus, detection of rubella virus in chorionic villus samples by RT-PCR may not always correctly predict fetal rubella virus infection. RT-PCR was successfully used for the diagnosis of congenitally acquired rubella in infancy. Rubella virus RNA was detected in cyropreserved or formalin-fixed lens aspirates obtained from infants in India with serologically confirmed congenital rubella but not in samples from controls with inherited cataract.
对一种逆转录巢式聚合酶链反应检测法(RT-PCR)进行了评估,以用于诊断子宫内及婴儿期先天性风疹感染。将RT-PCR与病毒分离法进行比较,以回顾性检测原发性风疹或风疹病毒再感染后终止妊娠所获得的胎盘和胎儿组织中的风疹病毒。85%的样本获得了一致结果;4个样本仅通过RT-PCR检测到风疹病毒RNA,2个样本仅通过病毒分离法检测到风疹病毒。还获取了样本用于先天性感染的产前诊断;通过RT-PCR在7个绒毛膜绒毛样本中的3个以及3个羊水样本中的1个检测到风疹病毒RNA,而仅在1个绒毛膜绒毛样本中分离到风疹病毒。为证明从绒毛膜绒毛样本中提取的RNA含有可扩增的RNA,采用巢式RT-PCR检测角蛋白mRNA。在2例胎儿未感染的病例中,在胎盘中检测到风疹病毒,而在1例胎儿感染的病例中,胎盘中未发现风疹病毒的证据。因此,通过RT-PCR检测绒毛膜绒毛样本中的风疹病毒可能并不总能正确预测胎儿风疹病毒感染。RT-PCR成功用于婴儿期先天性风疹感染的诊断。在印度从血清学确诊为先天性风疹的婴儿中获取的冷冻保存或福尔马林固定的晶状体吸出物中检测到风疹病毒RNA,但在遗传性白内障对照样本中未检测到。