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目的 :比较不同取样和提取方法对甲状腺结节细针穿刺(fine needle aspiration,FNA)标本提取RNA检测目的基因结果的影响。方法:以单独穿刺与穿刺针冲洗液2种方式收取FNA标本,以TRIzol法和Qiagen法2种方法提取RNA,利用反转录-聚合酶链反应(reverse transcription-polymerase chain reaction,RT-PCR)检测甲状腺细胞相关基因,比较不同方法对RNA质与量及RT-PCR结果的影响。结果:102例FNA标本均提取了RNA,RT-PCR结果显示,GAPDH的检出率达97.06%,TG为93.14%,其余依次为TPO、TSHr、NIS。在单独穿刺样本中,TRIzol法提取的RNA浓度及总量均大于Qiagen法,但差异未达统计学意义(P>0.05)。2种方法所提取RNA的吸光度值及PCR成功率也无统计学差异(P>0.05)。在冲洗液标本中亦未发现Qiagen法与TRIzol法在RNA浓度、总量、吸光度值及PCR成功率之间有显著差异(P>0.05)。在TRIzol组中,单独穿刺与穿刺针冲洗液2种取样方式对其RNA浓度、RNA总量、吸光度值以及PCR成功率的影响均无统计学意义(P>0.05)。在Qiagen组中,不同取样方式对其RNA浓度、RNA总量、OD值以及PCR成功率的影响亦无统计学意义(P>0.05)。结论:FNA标本可以成功提取RNA并进行基因检测。单独穿刺、穿刺针冲洗液2种取样方式以及TRIzol法、Qiagen法2种RNA提取方法均可用于FNA标本RNA的提取。
Objective: To compare the effects of different sampling and extraction methods on the results of RNA extraction from fine needle aspiration (FNA) specimens of thyroid nodules. Methods: FNA specimens were collected by two ways of puncturing and puncturing needle separately. RNA was extracted by two methods of TRIzol and Qiagen. Reverse transcription-polymerase chain reaction (RT-PCR) Detection of thyroid cells related genes, compare different methods of RNA quality and quantity and RT-PCR results. Results: RNA was extracted from 102 FNA specimens. The results of RT-PCR showed that the detection rate of GAPDH was 97.06% and the TG was 93.14%. The rest were TPO, TSHr and NIS. In single puncture samples, the concentration and total RNA extracted by TRIzol method were greater than that of Qiagen method, but the difference was not statistically significant (P> 0.05). There was no significant difference in absorbance value and PCR success rate between the two methods (P> 0.05). There was no significant difference (P> 0.05) between the RNA concentration, the total amount, the absorbance value and the PCR success rate between the Qiagen method and the TRIzol method in the washing solution samples. In the TRIzol group, there were no significant differences in RNA concentration, RNA content, absorbance value and PCR success rate between the two sampling methods of puncture and puncture needle wash alone (P> 0.05). In Qiagen group, the effect of different sampling methods on RNA concentration, total RNA amount, OD value and PCR success rate was also not statistically significant (P> 0.05). Conclusion: FNA samples can be successfully extracted RNA and genetic testing. Two kinds of sampling methods, such as puncture and puncture needle wash alone, and TRIzol method and Qiagen method, can be used to extract RNA from FNA specimens.