间接法原位PCR在cHL IgH基因重排研究中的应用

Application of indirect in situ PCR in detection of IgH gene rearrangement

目的 :探索在石蜡切片上进行间接法原位PCR的可行性及利用产物探针检测IgH克隆性基因重排的意义。 方法 :选取获得IgH克隆性基因重排的经典型霍奇金淋巴瘤 (cHL) ,纯化产物合成探针 ,进行间接法原位PCR。 结果 :在H/RS细胞和周围背景细胞核内均发现棕褐色杂交颗粒 ,利用产物探针互测时也能出现杂交颗粒。结论 :在石蜡切片上进行间接法原位PCR是可行和有效的 ,但产物探针的精确性仍有待进一步证实。

Purpose To explore the feasibility to proceed in situ detection of DNA after PCR in paraffin section and its significance to detect the IgH gene rearrangement. Methods Dig labeled probes were synthesized by use of purified IgH PCR products, with which the authors proceeded in situ detection of DNA after PCR in classic Hodgkin's lymphoma. Results The method of in situ detection of DNA after PCR was established in paraffin embedded section and found brown staining particles not only in H/RS cells' nuclei but also in the surrounding background cells'. So did that when complementary probes were used. Conclusions It is feasible and valid to proceed in situ detection of DNA after PCR in paraffin section. But the accuracy of PCR product probe is limited.