摘要
目的探讨应用于乙肝携带者卵巢颗粒细胞涂片原位杂交技术。方法选择6例乙肝携带者颗粒细胞涂片原位杂交,每例涂片12张,分别进行蛋白酶K浓度(1μg、2μg、3μg)、消化时间(1min、3min、5min)、探针浓度(1∶10、1∶20、1∶30)及显色时间(10min、20min、30min)优化。结果 3例颗粒细胞核均检测到HBVDNA阳性信号。最佳反应条件为,蛋白酶K浓度为2μg,消化时间为3min,探针∶预杂交液比为1∶20,显色时间为20min。结论蛋白酶K浓度、消化时间、探针浓度及显色时间是原位杂交中关键性步骤,该技术可用于颗粒细胞涂片的研究。
Objective To explore an in situ hybridization method which could be used in human granulose cell smear of HBV carriers to detect HBV DNA.Methods 6 granulose cell smear cases of hepatitis B carrier [HBsAg（＋）,HBeAg（＋）,HBcAb（＋）] were detected.Each has 12 slides.The working conditions of proteinase K concentration,digestion time,probe concentration and color reaction time were optimazed.Results HBV DNA were detected in 3 cases of granulose cell smears.The best working conditions were as follows：the condition of proteinase K was 2 μg,digestion time was 3 min,the ratio of probe concentration was 1：20,and colour reaction time was 20 min.Conclusions The proteinase K concentration,digestion time,probe concentration and chromogenic are crucial steps of in situ hybridization technique.The technology can be applied to study human granulose cell smear.
出处
《北京医学》
CAS
2010年第9期742-743,共2页
Beijing Medical Journal
关键词
颗粒细胞
原位杂交
乙型肝炎病毒
Granulose cell In situ hybridization Hepatitis B virus
