As an immediate-early protein of herpes simplex virus, infected-cell polypeptide 0 (ICP0) exhibits complicated interactions with host cells, and its regulatory function on gene expression is of great importance. Since...As an immediate-early protein of herpes simplex virus, infected-cell polypeptide 0 (ICP0) exhibits complicated interactions with host cells, and its regulatory function on gene expression is of great importance. Since the ICP0 encoding sequence contains many rare codons which are absent in E.coli, and ICP0 is highly unstable in prokaryotic cells, expression of entire ICP0 in prokaryotic cells has never been reported. In order to further investigate the function of ICP0, a recombinant plasmid was constructed by subcloning a cDNA fragment encoding an amino-terminal of 105 residues of the ICP0 protein into pGEX-5x-1 vector. The resulting GST-105 fusion antigen peptide was expressed with high efficiency in E.coli. Antibodies prepared after the immunization of mice with purified fusion protein can recognize not only the denatured ICP0 protein, but also the native ICP0 protein with normal biological conformation.展开更多
目的 比较常用的3种检测单纯疱疹性角膜炎(HSK)辅助诊断方法的敏感性和临床实用性。方法6只新西兰白兔双眼接种1×105PFU单纯疱疹病毒I型(HSV-I)McKrea株。病毒接种后第8天应用无环鸟苷眼膏每2 h 1次,共4次。另外2只兔中1只兔双眼...目的 比较常用的3种检测单纯疱疹性角膜炎(HSK)辅助诊断方法的敏感性和临床实用性。方法6只新西兰白兔双眼接种1×105PFU单纯疱疹病毒I型(HSV-I)McKrea株。病毒接种后第8天应用无环鸟苷眼膏每2 h 1次,共4次。另外2只兔中1只兔双眼在角膜上划痕但不接种病毒,另1只兔双眼角膜接种曲霉菌作为阴性对照。在病毒接种后的3、9、21天,利用病毒分离、多聚酶链反应(PCR)和蛋白吸附膜快速单纯疱疹病毒I型抗原检测方法进行检测。结果 接种后第2天全部角膜出现典型的HSK点状或树枝状浸润,第5天出现地图状溃疡,第21天仍有2只角膜有持续性感染。抗病毒治疗1天后无明显好转。2只兔于接种后14和16天死亡。病毒分离方法只有接种后3天的泪液培养阳性,第9、21天的标本检测均为阴性。有11/12、11/12和6/8只角膜在接种后的第3、9、21天用PCR方法检测到了HSV-I DNA。蛋白吸附膜抗原检测在接种后的第3、9、21天的阳性率分别为12/12、12/12和3/8。结论 蛋白吸附膜快速HSV—I检测是一种快速、简便、经济的方法。其阳性结果与临床表现相吻合。展开更多
目的猴B病毒(monkey B virus,BV),也称猴疱疹病毒I型(Cercopithecine herpesvirus 1),是重要的人兽共患病原。根据国家标准,猴B病毒作为抗原检测抗体,但由于生物安全问题,抗原制备受到很大限制,因此使用替代抗原进行抗体血清学检测并进...目的猴B病毒(monkey B virus,BV),也称猴疱疹病毒I型(Cercopithecine herpesvirus 1),是重要的人兽共患病原。根据国家标准,猴B病毒作为抗原检测抗体,但由于生物安全问题,抗原制备受到很大限制,因此使用替代抗原进行抗体血清学检测并进行比对验证。方法应用2种ELISA方法(抗原分别为BV和HVP2)和1种免疫酶法EIA方法(抗原为HSV-1)对本实验室135份送检恒河猴血液样品进行筛查,对阳性及可疑样品再以免疫荧光法IFA和Western blot方法(抗原为HSV-1)以及以HSV-1 g C1纯化糖蛋白为抗原的免疫印迹方法验证。结果HVP2-ELISA、BV-ELISA和HSV-1-EIA阳性检出率分别为32.6%、37.8%和34.8%;3种方法检测结果一致的样品占91.1%(123/135),阳性结果可被IFA和WB确证;可疑样品12份,33.3%(4/12)的样品经验证检验为阳性。结论与BV抗原相比,替代抗原HSV-1的敏感性和特异性较HVP2更为接近;阳性样品及可疑样品的确证检验应使用多种方法,避免漏检。展开更多
基金National natural science foundation items (30570081 and 30670094)
文摘As an immediate-early protein of herpes simplex virus, infected-cell polypeptide 0 (ICP0) exhibits complicated interactions with host cells, and its regulatory function on gene expression is of great importance. Since the ICP0 encoding sequence contains many rare codons which are absent in E.coli, and ICP0 is highly unstable in prokaryotic cells, expression of entire ICP0 in prokaryotic cells has never been reported. In order to further investigate the function of ICP0, a recombinant plasmid was constructed by subcloning a cDNA fragment encoding an amino-terminal of 105 residues of the ICP0 protein into pGEX-5x-1 vector. The resulting GST-105 fusion antigen peptide was expressed with high efficiency in E.coli. Antibodies prepared after the immunization of mice with purified fusion protein can recognize not only the denatured ICP0 protein, but also the native ICP0 protein with normal biological conformation.
文摘目的猴B病毒(monkey B virus,BV),也称猴疱疹病毒I型(Cercopithecine herpesvirus 1),是重要的人兽共患病原。根据国家标准,猴B病毒作为抗原检测抗体,但由于生物安全问题,抗原制备受到很大限制,因此使用替代抗原进行抗体血清学检测并进行比对验证。方法应用2种ELISA方法(抗原分别为BV和HVP2)和1种免疫酶法EIA方法(抗原为HSV-1)对本实验室135份送检恒河猴血液样品进行筛查,对阳性及可疑样品再以免疫荧光法IFA和Western blot方法(抗原为HSV-1)以及以HSV-1 g C1纯化糖蛋白为抗原的免疫印迹方法验证。结果HVP2-ELISA、BV-ELISA和HSV-1-EIA阳性检出率分别为32.6%、37.8%和34.8%;3种方法检测结果一致的样品占91.1%(123/135),阳性结果可被IFA和WB确证;可疑样品12份,33.3%(4/12)的样品经验证检验为阳性。结论与BV抗原相比,替代抗原HSV-1的敏感性和特异性较HVP2更为接近;阳性样品及可疑样品的确证检验应使用多种方法,避免漏检。
