原位杂交检测微小病毒B19方法的建立及临床应用

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９ＣＨＤ）心脏组织细胞的定位分布。方法 以Ｂ１９特异的衣壳蛋白ＶＰＩ ＤＮＡ内１１１２ｂｐ片段为模板，采用随机引物标记探针法，建立了ＩＳＨ检测Ｂ１９ ＤＮＡ浓度于０．１ｂｇ／μ１以上呈阳性。在６６例先天性心脏病心脏组织中，检测至Ｂ１９ ＤＮＡ阳性７例，并发现Ｂ１９ ＤＮＡ阳性信号主要定位于心肌细胞核内，３８例对照组健康心肌

Establishment ofin situ hybridization technique for detection of parvovirus B19 and its clinical application

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 1112 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/microl, 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. CONCLUSIONS: We have established ISH to detect B19 DNA that has high sensitivity (0. 1 pg/microl) and specificity and have found that B19 DNA is located the nucleus of cardiac cell.