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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,并确定B19DNA定位于心肌组织的心肌细胞核内
Objective To establish an in situ hybridization (ISH) method for the detection of human parvovirus B19 and determine its localization in cardiac tissue of patients with congenital heart disease (CHD). Methods The 1112bp fragment of B19 specific capsid protein VP1DNA was used as a template to detect B19 DNA by ISH. Results The positive results of dot blot hybridization showed that B19 DNA was positive, and negative for CMV, HSV and TOXDNA. The quantitative analysis of B19 DNA was positive at more than 0.1 pg / μl. In 66 cases of congenital heart disease, 7 cases of positive B19DNA were detected. The positive signal of B19DNA was mainly located in the nucleus of myocardial cells, and 38 cases of healthy controls were negative. Conclusions The established method of ISH for detection of B19 in heart tissue of congenital heart disease has good specificity and sensitivity with a sensitivity of 0.1 pg / μl, and confirms that B19 DNA is localized in the myocardial nucleus of myocardium