Puchhammer-Stoeckl E, Heinz F X, Kunz C
Institute of Virology, University of Vienna, Austria.
J Virol Methods. 1993 Jul;43(2):257-66. doi: 10.1016/0166-0934(93)90081-2.
The efficiencies of 4 different hybridization assays, using probes labeled with 32P, digoxigenin (DIG) and biotin, were compared for the detection of herpes simplex virus (HSV) DNA amplified from cerebrospinal fluid during herpes simplex encephalitis (HSE). The biotinylated probe was shown to provide a clearly lower sensitivity than the radioactive hybridization system, regardless of whether the biotin-labeled probe was detected by a colorimetric or by a photobiotin assay. In contrast, the DIG system, which includes a luminescent detection step was shown to be equivalent to the 32P system, at least at lower template concentrations. Although generally higher background signals were observed with the DIG assay the analysis of 73 CSF samples yielded identical results with the 32P and the DIG test systems in all cases, identifying concordantly an HSE in 6 patients. Since the time necessary for the performance of both assays is similar the DIG system seems to be a powerful alternative to the radioactive detection of amplified HSV DNA.
为检测单纯疱疹性脑炎（HSE）期间从脑脊液中扩增出的单纯疱疹病毒（HSV）DNA，比较了使用32P、地高辛配基（DIG）和生物素标记探针的4种不同杂交检测方法的效率。结果显示，无论生物素标记探针是通过比色法还是光生物素检测法进行检测，生物素化探针的灵敏度均明显低于放射性杂交系统。相比之下，包括发光检测步骤的DIG系统显示与32P系统相当，至少在较低模板浓度下如此。虽然DIG检测通常观察到较高的背景信号，但对73份脑脊液样本的分析在所有情况下32P和DIG检测系统均产生相同结果，一致鉴定出6例患者患有HSE。由于两种检测方法所需时间相似，DIG系统似乎是扩增HSV DNA放射性检测的有力替代方法。
