The establishment and application of internal quality control system for real-time quantitative PCR detection of BCR-ABL (P210) transcript levels
10.3760/cma.j.issn.0253-2727.2016.09.014
- VernacularTitle:实时定量PCR检测BCR-ABL(P210)转录本水平室内长期质控体系的建立及应用
- Author:
Chaoqin ZHONG
1
;
Na HE
;
Mingqiang HUA
;
Xiaodong WEI
;
Daoxin MA
;
Chunyan JI
Author Information
1. 山东大学齐鲁医院血液科
- Keywords:
Real-time polymerase chain reaction;
Fusion proteins,BCR-ABL;
Internal quality control;
Slope rate;
Intercept
- From:
Chinese Journal of Hematology
2016;37(9):800-806
- CountryChina
- Language:Chinese
-
Abstract:
Objective To set internal quality control system of BCR-ABL(P210) transcript levels for real-time quantitative PCR (RQ-PCR).Methods Using K562 cells and HL-60 cells,we prepared high-and low-level BCR-ABL internal quality control substance.The BCR-ABL (P210) transcript levels of internal quality control substance have been determined for 184 times together with clinical samples from August 2013 to October 2015.The slope rate,intercept and correlation coefficient of standard curve were calculated according to different reagent lots (lots number 20130303,20131212,20140411 and 20150327 are called R1、R2、R3 and R4 for short respectively),and the detection results of quality control substance were calculated according to different reagent lots and quality control substance lots (lots number 20130725,20140611 are called Q 1、Q2 for short respectively).Then the results were analyzed by Levey-Jennings quality control chart combined with Westgard multi-rules theory.Results ①We analyzed the slope rate and intercept of standard curve.Fifty-three times of the R1 reagent detection,80 times of the R3 reagent detection and 14 times of the R4 reagent detection were all under control.For 37 times detection of R2 reagent,the slope rate was out of control for 6 times.It was lower than-x-s for the 2-8 tests and upper the average for the 12-37 tests.The intercept was out of control for 9 times,upper the-x+s for the 1-8 tests and lower the average for the 12-37 tests.②According to the detection results of quality control substance,for Q1 quality control substance,49 tests by R1 reagent were under control,and 1 out of 23 tests by R2 reagent was out of control.For Q2 quality control substance,14 tests by R2 reagent detection,72 tests by R3 reagent detection and 14 tests by R4 reagent were all under control.Conclusion The preparation of high-and low-level quality control substance using K562 and HL-60 cells was convenient and the detection results were reliable and stable.The application of quality control substance combined with slope rate and intercept in the internal quality control may contribute to quality assurance forquantitative detection of BCR-ABL (P210) transcript levels.
