An interlaboratory comparison study on the detection of RUNX1-RUNX1T1 fusion transcript levels and WT1 transcript levels.

Ya Zhen Qin; Li Wen Zhu; Shang Lin; Su Xia Geng; Sheng Wei Liu; Hui Cheng; Cheng Ye WU; Min Xiao; Xiao Qing LI; Rui Ping HU; Li Li Wang; Hai Yan Liu; Dao Xin MA; Tao Guan; Yuan Xin YE; Ting Niu; Jian Nong Cen; Li Sha LU; Li Sun; Tong Hua Yang; Yun Gui Wang; Tao LI; Yue Wang; Qing Hua LI; Xiao Su Zhao; Ling Di LI; Wen Min Chen; Ling Yu Long; Xiao Jun Huang

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An interlaboratory comparison study on the detection of RUNX1-RUNX1T1 fusion transcript levels and WT1 transcript levels.

Author: Ya Zhen QIN ¹ ; Li Wen ZHU ² ; Shang LIN ³ ; Su Xia GENG ⁴ ; Sheng Wei LIU ⁵ ; Hui CHENG ⁶ ; Cheng Ye WU ⁷ ; Min XIAO ⁸ ; Xiao Qing LI ⁹ ; Rui Ping HU ¹⁰ ; Li Li WANG ¹¹ ; Hai Yan LIU ¹² ; Dao Xin MA ¹³ ; Tao GUAN ¹⁴ ; Yuan Xin YE ¹⁵ ; Ting NIU ¹⁶ ; Jian Nong CEN ¹⁷ ; Li Sha LU ¹⁸ ; Li SUN ¹⁹ ; Tong Hua YANG ²⁰ ; Yun Gui WANG ²¹ ; Tao LI ²² ; Yue WANG ²³ ; Qing Hua LI ²⁴ ; Xiao Su ZHAO ¹ ; Ling Di LI ¹ ; Wen Min CHEN ¹ ; Ling Yu LONG ¹ ; Xiao Jun HUANG ¹
Author Information

1. Peking University People's Hospital, Peking University Institute of Hematology, National Clinical Research Center for Hematologic Disease, Beijing 100044, China.
2. Beijing Hightrust Diagnostics Co., Ltd, Beijing 100176, China.
3. Department of Hematology, The First Affiliated Hospital of Dalian Medical University, Dalian 116011, China.
4. Department of Hematology, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou 510080, China.
5. Harbin Institute of Hematology and Oncology, Harbin 150010, China.
6. Changhai Hospital, The Second Military Medical University, Shanghai 200433, China.
7. Institute of Hematology, Henan Provincial People's Hospital, Zhengzhou 450003, China.
8. Department of Hematology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030.
9. Center for Stem Cell Research and Application, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022.
10. Department of Hematology, Bethune First Affiliated Hospital of Jilin University, Changchun 130021.
11. Department of Hematology, Chinese PLA General Hospital, Beijing 100853, China.
12. Department of Hematology, Affiliated Hospital of Nantong University, Nantong 226001, China.
13. Department of Hematology, Qilu Hospital of Shandong University, Jinan 250012, China.
14. Department of Hematology, Shanxi Provincial Cancer Hospital, Taiyuan 030000, China.
15. Department of Laboratory Medicine, West China Hospital of Sichuan University, Chengdu 610041, China.
16. Department of Hematology, West China Hospital of Sichuan University, Chengdu 610041, China.
17. The First Affiliated Hospital of Soochow University, Jiangsu Institute of Hematology, National Clinical Research Center for Hematologic Disease, Suzhou 215006, China.
18. Tianjin Sino-us Diagnostics Co., Ltd, Tianjin 301617, China.
19. Wuhan Kindstar Diagnostics Co., Ltd, Wuhan 430075, China.
20. Department of Hematology, The First People's Hospital of Yunnan Province, Kunming 650034, China.
21. Department of Hematology, The First Affiliated Hospital, Zhejiang University, Hangzhou 310003, China.
22. Department of Hematology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China.
23. The First Hospital of China Medical University, Shenyang 110001, China.
24. Institute of Hematology and Blood Disease Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, National Clinical Research Center for Hematologic Disease, Tianjin 300020, China.
Publication Type:Journal Article
Keywords: Fusion protein, RUNX1-RUNX1T1; Interlaboratory comparison; Real-time quantitative PCR; WT1
MeSH: China; Core Binding Factor Alpha 2 Subunit; Humans; Leukemia, Myeloid, Acute; RUNX1 Translocation Partner 1 Protein; Real-Time Polymerase Chain Reaction; Transcription, Genetic; WT1 Proteins
From: Chinese Journal of Hematology 2019;40(11):889-894
CountryChina
Language:Chinese
Abstract: Objective: To investigate the current status and real performance of the detection of RUNX1-RUNX1T1 fusion transcript levels and WT1 transcript levels in China through interlaboratory comparison. Methods: Peking University People's Hospital (PKUPH) prepared the samples for comparison. That is, the fresh RUNX1-RUNX1T1 positive (+) bone morrow nucleated cells were serially diluted with RUNX1-RUNX1T1 negative (-) nucleated cells from different patients. Totally 23 sets with 14 different samples per set were prepared. TRIzol reagent was added in each tube and thoroughly mixed with cells for homogenization. Each laboratory simultaneously tested RUNX1-RUNX1T1 and WT1 transcript levels of one set of samples by real-time quantitative PCR method. All transcript levels were reported as the percentage of RUNX1-RUNX1T1 or WT1 transcript copies/ABL copies. Spearman correlation coefficient between the reported transcript levels of each participated laboratory and those of PKUPH was calculated. Results: ①RUNX1-RUNX1T1 comparison: 9 samples were (+) and 5 were (-) , the false negative and positive rates of the 20 participated laboratories were 0 (0/180) and 5% (5/100) , respectively. The reported transcript levels of all 9 positive samples were different among laboratories. The median reported transcript levels of 9 positive samples were from 0.060% to 176.7%, which covered 3.5-log. The ratios of each sample's highest to the lowest reported transcript levels were from 5.5 to 12.3 (one result which obviously deviated from other laboratories' results was not included) , 85% (17/20) of the laboratories had correlation coefficient ≥0.98. ②WT1 comparison: The median reported transcript levels of all 14 samples were from 0.17% to 67.6%, which covered 2.6-log. The ratios of each sample's highest to the lowest reported transcript levels were from 5.3-13.7, 62% (13/21) of the laboratories had correlation coefficient ≥0.98. ③ The relative relationship of the reported RUNX1-RUNX1T1 transcript levels between the participants and PKUPH was not always consistent with that of WT1 transcript levels. Both RUNX1-RUNX1T1 and WT1 transcript levels from 2 and 7 laboratories were individually lower than and higher than those of PKUPH, whereas for the rest 11 laboratories, one transcript level was higher than and the other was lower than that of PKUPH. Conclusion: The reported RUNX1-RUNX1T1 and WT1 transcript levels were different among laboratories for the same sample. Most of the participated laboratories reported highly consistent result with that of PKUPH. The relationship between laboratories of the different transcript levels may not be the same.