摘要
目的 建立原位杂交 (ISH)方法检测人微小病毒B19,并确定其在先天性心脏病 (CHD)心脏组织细胞的定位分布。方法 以B19特异的衣壳蛋白VP1DNA内 1112bp片段为模板 ,采用随机引物标记探针法 ,建立了ISH检测B19DNA。结果 通过斑点杂交定性分析显示B19DNA呈阳性 ,CMV、HSV和TOXDNA均呈阴性 ;定量分析B19DNA浓度于 0 .1pg/ μl以上呈阳性。在 6 6例先天性心脏病心脏组织中 ,检测到B19DNA阳性 7例 ,并发现B19DNA阳性信号主要定位于心肌细胞核内 ,38例对照组健康心肌中均阴性。结论 建立的ISH检测先天性心脏病心脏组织B19的方法有良好的特异性及敏感性 ,敏感性达 0 .1pg/ μl。
Objective To establish In situ hybridization(ISH) technique to detect parvovirus B19 and to know the viral location in cardiac tissue of congenital heart diseases(CHD). Methods ISH method was established with randomly primed,degoxigenin labeled B19 DNA probe of 1 112 bp long corresponding to parvovirus B19 capsid protein VP1 genome.Results Dot blot hybridization showed that B19 DNA was positive with the sensitivity of 0.1 pg/μl, while CMV、HSV、TOX DNA were all negative. 7 out of 66 CHD cardiac tissues were B19 DNA positive by using ISH, while in 38 cases of control group were all negative. B19 DNA positive signal was found in the nucleus of cardiac cell.Conclusion We have established ISH to detect B19 DNA that has high sensitivity(0.1 pg/μl)and specificity and have found that B19 DNA is located the nucleus of cardiac cell.
出处
《中华实验和临床病毒学杂志》
CAS
CSCD
2000年第3期268-270,共3页
Chinese Journal of Experimental and Clinical Virology
基金
国家计生委基金助课题 !(9736 )
