Feddersen Søren, Bastholt Lars, Pedersen Susanne M
1 Department of Clinical Biochemistry and Pharmacology, Odense University Hospital, Odense, Denmark.
2 Department of Clinical Research, University of Southern Denmark, Odense, Denmark.
Ann Clin Biochem. 2017 Sep;54(5):558-566. doi: 10.1177/0004563216671538. Epub 2016 Sep 13.
Background The clinical utility of serum thyroglobulin in the follow-up of patients with differentiated thyroid carcinoma may be compromised by the presence of endogenous antithyroglobulin antibodies. To prevent interference by antithyroglobulin antibodies several groups have developed real-time PCR-based assays for quantification of blood thyroglobulin mRNA levels. For accurate quantification of thyroglobulin mRNA in blood preanalytical factors must be recognized and controlled. In this study, we evaluate the effect of different blood RNA stabilizing systems - the Tempus Blood RNA system and the PAXgene Blood RNA system - and storage time on RNA yield and quality, and thyroglobulin mRNA stability. Methods Blood samples from 11 patients previously treated for differentiated thyroid carcinoma were collected in K-EDTA, Tempus and PAXgene tubes and maintained at room temperature. RNA was isolated following storage for 0 and 72 h, and RNA yield, integrity and purity was determined. Thyroglobulin, GAPDH and ACTB mRNA levels were quantified by semi-quantitative real-time PCR. Results The RNA yield was significantly higher for blood collected in Tempus tubes compared with PAXgene tubes following storage for 72 h at room temperature ( P = 0.0011). High-quality RNA could be extracted from blood collected in PAXgene and Tempus tubes. Blood collected in K-EDTA tubes, but not in PAXgene and Tempus tubes, showed significant changes in thyroglobulin mRNA levels following storage for 72 h at room temperature ( P = 0.0263). Conclusions Stabilization of blood in PAXgene and Tempus tubes enables storage at room temperature for up to 72 h, without compromising thyroglobulin mRNA levels.
内源性抗甲状腺球蛋白抗体的存在可能会影响血清甲状腺球蛋白在分化型甲状腺癌患者随访中的临床应用价值。为防止抗甲状腺球蛋白抗体的干扰，多个研究小组已开发出基于实时PCR的检测方法，用于定量血液中甲状腺球蛋白mRNA水平。为了准确量化血液中的甲状腺球蛋白mRNA，必须识别并控制分析前因素。在本研究中，我们评估了不同的血液RNA稳定系统——Tempus血液RNA系统和PAXgene血液RNA系统——以及储存时间对RNA产量和质量，以及甲状腺球蛋白mRNA稳定性的影响。方法：收集11例曾接受分化型甲状腺癌治疗患者的血液样本，分别置于K-EDTA管、Tempus管和PAXgene管中，并在室温下保存。分别于储存0小时和72小时后提取RNA，测定RNA产量、完整性和纯度。通过半定量实时PCR定量甲状腺球蛋白、甘油醛-3-磷酸脱氢酶（GAPDH）和β-肌动蛋白（ACTB）mRNA水平。结果：室温下储存72小时后，Tempus管收集的血液RNA产量显著高于PAXgene管收集的血液（P = 0.0011）。可以从PAXgene管和Tempus管收集的血液中提取高质量的RNA。K-EDTA管收集的血液在室温下储存72小时后甲状腺球蛋白mRNA水平发生显著变化，而PAXgene管和Tempus管收集的血液则未出现这种情况（P = 0.0263）。结论：PAXgene管和Tempus管可使血液在室温下稳定储存长达72小时，而不影响甲状腺球蛋白mRNA水平。
