The influence of peripheral blood sample storage and delivery on the quantitative detection result of BCR-ABL (P210) transcript levels
10.3760/cma.j.issn.0253-2727.2021.03.008
- VernacularTitle: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
- Author:
Mingqiang HUA
1
;
Na HE
;
Chaoqin ZHONG
;
Xinyu YANG
;
Jinting LIU
;
Ruiqing WANG
;
Fengjiao HAN
;
Chen ZHANG
;
Daoxin MA
Author Information
1. 山东大学齐鲁医院血液科，济南 250012
- Keywords:
Real-time quantitative polymerase chain reaction;
Fusion proteins, BCR-ABL;
Internal quality control;
Storage conditions;
Delivery
- From:
Chinese Journal of Hematology
2021;42(3):224-229
- CountryChina
- Language:Chinese
-
Abstract:
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 ℃; low temperature, 2-8 ℃) , 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 10N) the original data [BCR-ABL copy number, ABL copy number, and BCR-ABL (P210) transcript levels]. Results:①Agarose 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.
