Koskiniemi M, Mannonen L, Kallio A, Vaheri A
Haartman Institute, Department of Virology, University of Helsinki, Finland.
J Virol Methods. 1997 Jan;63(1-2):71-9. doi: 10.1016/s0166-0934(96)02116-7.
We modified and optimized a new microplate hybridization assay to detect the varciella-zoster virus (VZV) PCR product, and studied cerebrospinal fluid (CSF) samples of 287 patients with meningitis, encephalitis or other neurological diseases or symptoms. Specific antibodies to VZV and reference antigens were determined by enzyme immunoassay from serum and CSF, they were then compared with clinical findings and with the results obtained by VZV-PCR using different detection methods for VZV-specific amplified DNA. VZV DNA was found in the CSF of 25 patients using the microplate hybridization assay and chemiluminescence detection for amplified DNA. All 25 CSF samples were also positive in Southern blotting. Among the patients, 10 had chickenpox, 4 had shingles, and 11 had no rash at all. The detection rate of VZV-specific DNA by microplate hybridization was 30% higher than that obtained by conventional agarose gel electrophoresis. In most patients the diagnosis was confirmed by demonstrating specific intrathecal antibody production to VZV but not to other viruses. These results indicate the presence of VZV in the central nervous system (CNS) in many patients with chickenpox or shingles, and even in patients without a rash. The microplate hybridization assay based on chemiluminescence detection improves considerably the detection rate of the VZV-PCR product compared to agarose gel electrophoresis and will add to the list of recognized VZV infections in the CNS. It is especially useful in cases where there is no cutaneous manifestation.
我们改良并优化了一种新型微孔板杂交检测法，用于检测水痘-带状疱疹病毒（VZV）的PCR产物，并对287例患有脑膜炎、脑炎或其他神经疾病或症状的患者的脑脊液（CSF）样本进行了研究。通过酶免疫测定法从血清和脑脊液中测定VZV特异性抗体和参考抗原，然后将其与临床发现以及使用不同检测方法对VZV特异性扩增DNA进行VZV-PCR所获得的结果进行比较。使用微孔板杂交检测法和化学发光检测扩增DNA，在25例患者的脑脊液中发现了VZV DNA。所有25份脑脊液样本在Southern印迹法中也呈阳性。在这些患者中，10例有水痘，4例有带状疱疹，11例根本没有皮疹。微孔板杂交法检测VZV特异性DNA的检出率比传统琼脂糖凝胶电泳法高30%。在大多数患者中，通过证明针对VZV而非其他病毒产生特异性鞘内抗体来确诊。这些结果表明，许多水痘或带状疱疹患者，甚至没有皮疹的患者，中枢神经系统（CNS）中都存在VZV。与琼脂糖凝胶电泳相比，基于化学发光检测的微孔板杂交检测法显著提高了VZV-PCR产物的检出率，并将增加CNS中已确认的VZV感染病例。它在没有皮肤表现的病例中尤其有用。
