摘要
目的：研究在存档石蜡包埋皮肤组织标本中提取RNA及进行逆转录PCR的可行性和稳定性。方法：分别抽取1997年、1999～2001年、2004年3个时间段的存档标本各10例，经过蛋白酶K消化后用异硫氰酸-酸性苯酚法提取RNA，设计3对β-Actin不同产物长度的引物，其目的片段的长度分别为445bp、205bp和112bp，比较了以各特异性下游引物及随机引物进行逆转录PCR的结果。结果：随机引物的阳性率很低，产物长度为112bp时很稳定，经卡方检验与另外两个长度间有显著性差异，而各年龄组间无统计学差异。结论：在石蜡包埋组织中提取RNA并进行逆转录PCR，方法稳定，可进行大规模的回顾性研究。
Objective： To investigave the feasibility and stability of the technique that purification RNA from paraffin- embedded skin tissue and performance RT - PCR. Methods： Three groups archival tissues were studied, each group had 10 cases. The sections were digested with proteinase K and RNA were purified by the guanidine isothiocyante- phenol protocol. Three paired primers of β- Aetin were designed and the PCR products were 445 bp, 205 bp, 112 bp respectively. The efficiency of RT- PCR with antisonse primers and random primer were compared. Results： The efficiency was low if the reverse transcription step was primed with random primer. The result of RT- PCR was stable under the amplicon of 112 bp and the specific antisense primer was primed. There were significant differences as compared with the other two amplieon sizes and no significant difference among the stored tissues of different time. Conclusion： The archival cutaneous tissue could be utilized as a source for retraspective study on a large scale.
出处
《中国麻风皮肤病杂志》
2006年第1期20-22,共3页
China Journal of Leprosy and Skin Diseases
