卵裂球细胞乙型肝炎病毒FISH检测法的建立

目的:建立一种灵敏、特异的卵裂球细胞乙型肝炎病毒(HBV)感染的检测方法。方法:根据HBV基因上的保守序列设计并合成引物,应用PCR技术制备探针并进行生物素标记。废弃的胚胎卵裂球经激光打孔,活检2~4个卵裂球细胞,利用生物素标记的探针,应用荧光原位杂交技术(FISH)检测HBV感染情况。结果:在HBV的感染者的38例子代胚胎卵裂球细胞,11例呈现阳性信号,健康对照组未检测到阳性信号。结论:荧光原位杂交技术(FISH)可用于卵裂球细胞HBV感染的检测,在体外受精-胚胎移植(IVF-ET)实践中对降低HBV垂直传播有重要意义。

Establishment of a FISH Method for HBV Detection in Human Blastomere

Objective To establish a sensitive and specific method for the detection of HBV in human blastomere.Methods According to the specific sequence of HBV genes,the primers were designed and synthesized,DNA probe was prepared by polymerase chain reaction（PCR） amplification and labeled with biotin.Discarded embryos were drilled by laser,and 2～4 cells were taken for biopsy.Fluorescence in situ hybridization（FISH） was performed to detect the existence of HBV DNA in the blastomere by using the biotinylated probe.Results FISH detection showed that 11 cases of 38 embryos from patients with HBV infection were positive,while no positive signal was found in healthy controls.Conclusion FISH technique could be performed for the detection of HBV in human blastomere cells,and it has great significance to reduce vertical transmission of HBV in IVF-ET.