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本文从HSV-1(SM_(44)株)DNA的EcoRI酶切片段中筛选出F片段,以缺口翻译制备α-~(32)P标记的探针,在判定其特异性和敏感性的基础上,用F片段探针直接检测了分别来自疱疹性角膜炎患者和尸检三叉神经节的临床标本。结果表明,FH段探针能检测到少至10pg DNA,除与HSV-1、HSV-2DNA杂交外,不与HCMV、ADV、兔肾细胞及人白细胞DNA杂交。10例疱疹性角膜炎患者的临床标本均呈阳性,与血清学鉴定结果一致;7例尸检三叉神经节标本中6例呈阳性,Southern印迹杂交分析证明,HSV以两种形式即完整的或部分的核酸序列存在于人三叉神经节中。本文为HSV DNA特异性酶切片段探针应用于临床诊断和流行病学研究奠定了基础。
In this study, F fragment was screened from the EcoRI fragment of HSV-1 (SM_ (44)) DNA, and a-32P-labeled probe was prepared by nick translation. The specificity and sensitivity of the probe , The F fragment probe was used to directly detect clinical specimens from patients with herpes keratitis and autopsy trigeminal ganglion respectively. The results showed that FH probes could detect as few as 10 pg DNA and did not hybridize with HCMV, ADV, rabbit kidney cells and human leukocyte DNA except for hybridization with HSV-1 and HSV-2 DNA. The clinical specimens of 10 cases of herpes keratitis were positive, consistent with the results of serological tests; 7 of 7 autopsy trigeminal ganglion specimens were positive, Southern blot hybridization analysis showed that HSV in two forms that is complete or part Of the nucleic acid sequences present in human trigeminal ganglia. This article lays the foundation for clinical diagnosis and epidemiological studies of HSV DNA-specific fragment probes.