外周血样本的存储和运输对BCR-ABL（P210）转录本水平定量检测结果的影响

OBJECTIVE: To explore the influence of storage and delivery conditions of the peripheral blood samples from patients with chronic myeloid leukemia（CML）on the real-time quantitative PCR（RQ-PCR）detection of the BCR-ABL（P210）transcript levels. METHODS: The peripheral blood samples of 84 CML patients were collected. The same sample was divided into different groups according to storage time（0, 6, 12, 24, 48, and 72 h）, temperature（room temperature, 18–24 °C; low temperature, 2–8 °C）, and vibration conditions（3, 6, and 12 h）. RQ-PCR was used to detect BCR-ABL（P210）transcript levels of the different groups. This study logarithmically transformed（log(10)(N)）the original data［BCR-ABL copy number, ABL copy number, and BCR-ABL（P210）transcript levels］. RESULTS: ① Agarose gel electrophoresis showed significant RNA degradation of samples after storage for 48 and 72 h at room temperature. ②Among the overall samples, the BCR-ABL copy number of the samples stored at room temperature for 48 and 72 h was significantly lower than that of the samples stored at low temperature（P<0.05）. However, the BCR-ABL（P210）transcript levels had no significant difference between samples stored at low temperature and room temperature. ③No significant changes were noted in the BCR-ABL（P210）transcript levels at different storage times（6, 12, 24, 48, and 72 h）regardless of storage temperature（P>0.05）compared with that at baseline（0 h, −0.56±1.51）. ④ The BCR-ABL copy number of the overall sample only decreased significantly（P<0.05）at 48 h（2.93±1.59）and 72 h（2.79±1.42）compared with that at baseline（0 h, 3.35±1.60）when stored at room temperature. The ABL copy number in the overall sample decreased significantly at 48 and 72 h（whether low and room temperature; P<0.05）. However, no significant changes were noted in the BCR-ABL（P210）transcript levels after vibration for 3 h（−1.29±1.81）, 6 h（−1.24±1.72）, and 12 h（−1.18±1.68; P>0.05）compared with that at baseline（0 h, −0.60±1.37）. CONCLUSION: Sample storage time, storage temperature, and vibration can interfere with the results of BCR-ABL and ABL copy number but have no significant effect on the quantitative determination of BCR-ABL（P210）transcript levels. This study provides strong support for the feasibility of transregional transportation of peripheral blood samples from patients with CML.