van Doornum G J J, Guldemeester J, Osterhaus A D M E, Niesters H G M
Department of Virology, Erasmus MC, Rotterdam, The Netherlands.
J Clin Microbiol. 2003 Feb;41(2):576-80. doi: 10.1128/JCM.41.2.576-580.2003.
Procedures using real-time technique were developed to demonstrate the presence of herpes simplex virus type 1 (HSV-1) and HSV-2, varicella zoster virus (VZV), and cytomegalovirus (CMV) in miscellaneous clinical specimens. The assays were compared to rapid culture using centrifugation followed by detection with monoclonal antibodies. A total of 711 consecutive samples were collected from different patient groups. Throat swabs were obtained from transplant patients; dermal or oral specimens were collected from patients suspected for VZV or HSV infection. Genital specimens were taken from patients who attended the Clinic for Sexually Transmitted Diseases at the Dijkzigt Hospital Rotterdam presenting with symptoms of a primary genital ulcer. Nucleic acid extraction was carried out using a MagnaPure LC instrument. The amplification steps were performed on the ABI Prism 7700 sequence detection system. To monitor the process of extraction and amplification, a universal control consisting of seal herpesvirus type 1 (PhHV-1) was added to the clinical specimens. By culture 127 of 668 (19%) samples were positive for HSV-1, 72 of 668 (10.8%) specimens were positive for HSV-2, and 17 of 366 (4.6%) were positive for VZV. Using real-time amplification the numbers of positive specimens were 143 of 668 (21.4%), 97 of 668 (14.5%), and 27 of 366 (7.4%), respectively. Eighty-six specimens were tested for CMV, 12 (14.0%) were positive by culture, and 17 (19.8%) were positive by real-time PCR. The clinical data of the patients with discrepant results were reviewed thoroughly. In all cases the patients with only real-time PCR-positive results could be considered as truly infected. We concluded that the real-time amplification technique is suitable for the detection of human herpesvirus infection. It offers a semiquantitative and reliable assay with a quick result that is more sensitive than rapid culture, especially for the diagnosis of HSV-2 and VZV infections.
已开发出使用实时技术的程序，以证明单纯疱疹病毒1型（HSV-1）、HSV-2、水痘带状疱疹病毒（VZV）和巨细胞病毒（CMV）在各种临床标本中的存在。将这些检测方法与使用离心法随后用单克隆抗体进行检测的快速培养法进行比较。总共从不同患者组中收集了711份连续样本。从移植患者中获取咽拭子；从疑似VZV或HSV感染的患者中收集皮肤或口腔标本。生殖器标本取自鹿特丹迪克齐赫特医院性传播疾病诊所出现原发性生殖器溃疡症状的患者。使用MagnaPure LC仪器进行核酸提取。扩增步骤在ABI Prism 7700序列检测系统上进行。为了监测提取和扩增过程，将由海豹疱疹病毒1型（PhHV-1）组成的通用对照添加到临床标本中。通过培养，668份样本中有127份（19%）HSV-1呈阳性，668份标本中有72份（10.8%）HSV-2呈阳性，366份中有17份（4.6%）VZV呈阳性。使用实时扩增法，阳性标本数量分别为668份中的143份（21.4%）、668份中的97份（14.5%）和366份中的27份（7.4%）。对86份标本进行了CMV检测，12份（14.0%）培养呈阳性，17份（19.8%）实时PCR呈阳性。对结果不一致的患者的临床数据进行了全面审查。在所有情况下，仅实时PCR呈阳性结果的患者可被视为真正感染。我们得出结论，实时扩增技术适用于检测人类疱疹病毒感染。它提供了一种半定量且可靠的检测方法，结果快速，比快速培养更敏感，尤其适用于HSV-2和VZV感染的诊断。
