Sakrauski A, Weber B, Kessler H H, Pierer K, Doerr H W
Institut für Medizinische Virologie, Universitätskliniken Frankfurt, Germany.
J Virol Methods. 1994 Dec;50(1-3):175-84. doi: 10.1016/0166-0934(94)90174-0.
Rapid diagnosis of herpes simplex encephalitis (HSE) can only be achieved by the polymerase chain reaction (PCR). In order to carry out PCR under routine conditions, it is of great importance to establish an easy DNA extraction protocol and especially a rapid and sensitive DNA detection method. In the present study, two different solid phase hybridization assays (Gen-Eti-K-DNA Enzyme Immunoassay (DEIA), Sorin Biomedica, Italy and Enzymun-Test DNA detection, Boehringer Mannheim, Germany) were compared for detection of PCR amplified HSV DNA polymerase genome region, using standard primers, from cerebrospinal fluid (CSF) samples. 122 CSF samples obtained from patients suffering from encephalitis and hospitalized at the University Clinics of Frankfurt and Graz during the period January 1992 to July 1993 were tested. To ascertain the sensitivity of the hybridization assays, dilution series of a plasmid, encoding the amplified region of the polymerase gene, were investigated. The detection limit of the DEIA assay was one copy of the plasmid/microliter, and the lowest amount of DNA which could be detected by the Enzymun assay as well as Southern blot was 10 copies/microliter. 15 CSF samples obtained from patients with HSE were found positive by the three assays. Concordant results were also obtained with CSF samples from non-HSE patients. The results of this study show that new hybridization systems guarantee a fast and high-sensitive detection of amplified HSV DNA. HSV PCR in CSF can be carried out routinely by the combined use of rapid hybridization and a simple extraction procedure.
单纯疱疹性脑炎（HSE）的快速诊断只能通过聚合酶链反应（PCR）来实现。为了在常规条件下进行PCR，建立一种简便的DNA提取方案，尤其是一种快速且灵敏的DNA检测方法至关重要。在本研究中，比较了两种不同的固相杂交检测方法（意大利Sorin Biomedica公司的Gen-Eti-K-DNA酶免疫测定法（DEIA）和德国Boehringer Mannheim公司的Enzymun-Test DNA检测法），用于检测从脑脊液（CSF）样本中使用标准引物扩增的单纯疱疹病毒（HSV）DNA聚合酶基因组区域。对1992年1月至1993年7月期间在法兰克福和格拉茨大学诊所住院的脑炎患者的122份CSF样本进行了检测。为确定杂交检测方法的灵敏度，研究了编码聚合酶基因扩增区域的质粒的稀释系列。DEIA检测法的检测限为每微升一个质粒拷贝，Enzymun检测法以及Southern印迹法能够检测到的最低DNA量为每微升10个拷贝。三种检测方法均发现15份来自HSE患者的CSF样本呈阳性。非HSE患者的CSF样本也得到了一致的结果。本研究结果表明，新的杂交系统能够保证对扩增的HSV DNA进行快速且高灵敏度的检测。通过快速杂交和简单提取程序的联合使用，可常规进行CSF中的HSV PCR检测。
