Transmission of herpes simplex virus from father to neonate.
Transmission of herpes simplex virus from father to neonate.
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单纯疱疹病毒从父亲传播给新生儿。
DOI：
10.1016/s0022-3476(83)80710-0
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发表时间：
1983
期刊：
影响因子：
--
通讯作者：
Corey,L
中科院分区：
文献类型：
--
作者：
Yeager,AS;Ashley,RL;Corey,L
方法在人包皮成纤维细胞中分离病毒和HSV血清型。中和滴度是否如前所述进行？HSV毒株的分型。HSV病毒株最初使用间接荧光抗体方法分型。随后，使用Lonsdale描述的技术的改进进行限制性酶分析。将表现出细胞病变效应的二倍体成纤维细胞培养物的细胞和上清液的冷冻等分试样解冻，并在32 p存在下在低磷酸盐培养基（新英格兰Nuclear，Boston，MA）中重新接种到BHK-21细胞中。当存在4+细胞病变效应时，用十二烷基硫酸钠裂解细胞，用苯酚提取混合物，然后用乙醇沉淀核酸。将核酸混合物用核糖核酸酶A和T-1（Sigma Chemical，St. Louis，MO）处理并用乙醇再沉淀。限制性内切核酸酶Hpa 1、Bam H1、Kpn 1、Eco R1和Xhol购自Bethesda Research Laboratories（盖瑟斯堡，MD），并在供应商规定的条件下进行数字化。通过在水平的0.7%琼脂糖凝胶中以3.5V/cm电泳18小时来分离DNA片段。电泳后，将凝胶真空干燥并置于Kodak XS-1医用X射线胶片（Eastman Kodak，罗切斯特，NY）上进行放射自显影。
METHODSVirus isolation and serolo~ y~ HSV strains were isolated in human foreskin fibroblast cells. Neutralization titers were performed as previously described? Typing of HSV strains. HSV strains were initially typed using an indirect fluorescent antibody method. 7 Subsequently, restriction enzyme analysis was performed using a modification of the technique described by Lonsdale) Frozen aliquots of cells and supernatants of diploid fibroblast cultures that exhibited cytopathic effect were thawed and reinoculated into BHK-21 cells in low-phosphate media in the presence of 32p (New England Nuclear, Boston, MA). When 4+ cytopathic effect was present, the cells were lysed with sodium dodecylsulfate, the mixture extracted with phenol, and the nucleic acid then precipitated with ethanol. The nucleic acid mixture was treated with ribonuclease A and T-1 (Sigma Chemical, St. Louis, MO) and reprecipitated with ethanol. The restriction endonucleases Hpa 1, Bam H1, Kpn 1, Eco R1, and Xhol were purchased from Bethesda Research Laboratories (Gaithersburg, MD), and digestions were carried out under conditions specified by the supplier. The DNA fragments were separated by electrophoresis in horizontal 0.7% agarose gels run at 3.5 V/cm for 18 hours. After electrophoresis, the gels were dried under vacuum and placed on Kodak XS-1 medical x-ray film (Eastman Kodak, Rochester, NY) for autoradiography.