Ashshi Ahmed M, Klapper Paul E, Cooper Robert J
University Virology, Laboratory Medicine Academic Group, School of Medicine, The University of Manchester, Oxford Road, M13 9WL, Manchester, UK.
J Infect. 2003 Jul;47(1):59-64. doi: 10.1016/s0163-4453(03)00057-4.
To develop a sensitive multiplex PCR to detect HCMV, HHV6 and HHV7, to test this PCR on urine specimens sent to the virus diagnostic laboratory and on stored urine samples from HIV-positive patients and their HIV-negative partners and to compare the sensitivity of the multiplex PCR with the diagnostic laboratory's routine service for the detection of HCMV.
Primers specific for each of the three viruses were combined in a multiplex PCR that was then optimised for sensitivity. This PCR was applied prospectively to 413 unselected routine urine specimens over a 1 year period and retrospectively to 258 urine specimens from 63 HIV-positive patients and 10 HIV-negative partners.
In the prospective study, the multiplex PCR detected 40 specimens positive for HCMV alone, 10 for HHV6, 3 for HHV7 and 3 with a dual infection of HCMV and HHV6. The sensitivity for HCMV was 93.5% by multiplex PCR compared to 28.3% by culture. HHV6 DNA was detected in 6 neonates (2-21 days) and HHV7 DNA in 2 neonates (4 and 20 days). In the retrospective study of HIV patients, HCMV was the most commonly detected virus (55.6%) compared to HHV6 (7.9%) and HHV7 (4.8%).
. The multiplex PCR was significantly more sensitive than non-DNA based procedures for the detection of HCMV. Urine may be a useful non-invasive specimen for the detection of HHV6 and HHV7 and their presence in neonates suggest perinatal transmission or the possibility of in utero infection.
开发一种灵敏的多重PCR以检测人巨细胞病毒（HCMV）、人疱疹病毒6型（HHV6）和人疱疹病毒7型（HHV7），在送往病毒诊断实验室的尿液标本以及来自HIV阳性患者及其HIV阴性伴侣的储存尿液样本上测试该PCR，并将多重PCR的灵敏度与诊断实验室检测HCMV的常规服务进行比较。
将针对这三种病毒各自的特异性引物组合在一个多重PCR中，然后对其灵敏度进行优化。在1年的时间里，将该PCR前瞻性地应用于413份未经选择的常规尿液标本，并回顾性地应用于来自63名HIV阳性患者和10名HIV阴性伴侣的258份尿液标本。
在前瞻性研究中，多重PCR检测出仅HCMV阳性的标本40份、HHV6阳性的10份、HHV7阳性的3份以及HCMV和HHV6双重感染的3份。多重PCR检测HCMV的灵敏度为93.5%，而培养法为28.3%。在6名新生儿（2 - 21天）中检测到HHV6 DNA，在2名新生儿（4天和20天）中检测到HHV7 DNA。在对HIV患者的回顾性研究中，HCMV是最常检测到的病毒（55.6%），相比之下HHV6为（7.9%），HHV7为（4.8%）。
多重PCR在检测HCMV方面比非基于DNA的方法明显更灵敏。尿液可能是检测HHV6和HHV7的有用非侵入性标本，它们在新生儿中的存在提示围产期传播或子宫内感染的可能性。
