CAJ | 학술논문

目的建立一种能够对样本中的单纯疱疹病毒(Hsv)进行准确鉴定分型的方法,并从生殖器疱疹(GH)患者皮损的水疱液中分离出1株HSV-2.方法从1例GH患者病变部位采集样本,样本接种Vero-E6细胞,待出现细胞病变后收获病毒感染物,提取病毒感染物的基因组.根据特异性PCR对分离出的病毒进行分型鉴定.结果分离的毒株引起Vero-E6典型细胞病变,病变细胞圆缩、融合.型特异性PCR反应及l%琼脂糖凝胶电泳鉴定结果表明,扩增出的片段大小约为183 bp,与预期的HSV-2型特异性片段长度相符,且与数据库的比对结果显示相似性为100%.结论通过细胞分离培养和特异性PCR分型鉴定,能够实现对样本中的HSV的准确分型鉴定.
목적 건립일충능구대양본중적단순포진병독(Hsv)진행준학감정분형적방법,병종생식기포진(GH)환자피손적수포액중분리출1주HSV-2.방법 종1례GH환자병변부위채집양본,양본접충Vero-E6세포,대출현세포병변후수획병독감염물,제취병독감염물적기인조.근거특이성PCR대분리출적병독진행분형감정.결과 분리적독주인기Vero-E6전형세포병변,병변세포원축、융합.형특이성PCR반응급l%경지당응효전영감정결과표명,확증출적편단대소약위183 bp,여예기적HSV-2형특이성편단장도상부,차여수거고적비대결과현시상사성위100%.결론 통과세포분리배양화특이성PCR분형감정,능구실현대양본중적HSV적준학분형감정.
Objective To develop a method of detecting the herpes simplex vires(HSV)genotype in clinical samples and isolate a strain of HSV-2 from the blister fluid in lesional skin of patients with genitalis herpes.Method Swab samples were collected from genital lesions of a genital hevpes patient and placed in transport media.Samples were inoculated Vero-E6 cells.After the appearance of the cytotoxicity,the infection mixtures were collected,and subjected to genomic DNA extraction.Based on the type-specific PCR,the genotype was detected.Results The isolated virus strain made the typical cytopathologic in Vero-E6 cells,and the cytopathologic cells contracted roundly andmerged.The agar gel electrophoresis assay showed that a DNA fragment of about 183 bp was successfully amplified,which matched with the length of expected specific segment of HSV-2.The sequence was compared with other known sequences,in Genebank by using BLAST.The results showed that the hornology was 100% with HSV-2.Conclusions This study showsthat a strain of HSV-2 could be successfully isolated based on virus isolation test and type-specific PCR assay.