Detection of 22 common leukemic fusion genes using a single-step multiplex qRT-PCR-based assay
Abstract Background Fusion genes generated from chromosomal translocation play an important role in hematological malignancies. Detection of fusion genes currently employ use of either conventional RT-PCR methods or fluorescent in situ hybridization (FISH), where both methods involve tedious methodo...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2017-07-01
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| Series: | Diagnostic Pathology |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s13000-017-0634-3 |
| _version_ | 1828195985672634368 |
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| author | Xiaodong Lyu Xianwei Wang Lina Zhang Zhenzhu Chen Yu Zhao Jieying Hu Ruihua Fan Yongping Song |
| author_facet | Xiaodong Lyu Xianwei Wang Lina Zhang Zhenzhu Chen Yu Zhao Jieying Hu Ruihua Fan Yongping Song |
| author_sort | Xiaodong Lyu |
| collection | DOAJ |
| description | Abstract Background Fusion genes generated from chromosomal translocation play an important role in hematological malignancies. Detection of fusion genes currently employ use of either conventional RT-PCR methods or fluorescent in situ hybridization (FISH), where both methods involve tedious methodologies and require prior characterization of chromosomal translocation events as determined by cytogenetic analysis. In this study, we describe a real-time quantitative reverse transcription PCR (qRT-PCR)-based multi-fusion gene screening method with the capacity to detect 22 fusion genes commonly found in leukemia. This method does not require pre-characterization of gene translocation events, thereby facilitating immediate diagnosis and therapeutic management. Methods We performed fluorescent qRT-PCR (F-qRT-PCR) using a commercially-available multi-fusion gene detection kit on a patient cohort of 345 individuals comprising 108 cases diagnosed with acute myeloid leukemia (AML) for initial evaluation; remaining patients within the cohort were assayed for confirmatory diagnosis. Results obtained by F-qRT-PCR were compared alongside patient analysis by cytogenetic characterization. Results Gene translocations detected by F-qRT-PCR in AML cases were diagnosed in 69.4% of the patient cohort, which was comparatively similar to 68.5% as diagnosed by cytogenetic analysis, thereby demonstrating 99.1% concordance. Overall gene fusion was detected in 53.7% of the overall patient population by F-qRT-PCR, 52.9% by cytogenetic prediction in leukemia, and 9.1% in non-leukemia patients by both methods. The overall concordance rate was calculated to be 99.0%. Fusion genes were detected by F-qRT-PCR in 97.3% of patients with CML, followed by 69.4% with AML, 33.3% with acute lymphoblastic leukemia (ALL), 9.1% with myelodysplastic syndromes (MDS), and 0% with chronic lymphocytic leukemia (CLL). Conclusions We describe the use of a F-qRT-PCR-based multi-fusion gene screening method as an efficient one-step diagnostic procedure as an effective alternative to lengthy conventional diagnostic procedures requiring both cytogenetic analysis followed by targeted quantitative reverse transcription (qRT-PCR) methods, thus allowing timely patient management. |
| first_indexed | 2024-04-12T09:53:36Z |
| format | Article |
| id | doaj.art-ac8440886d944b0f85b02f949433d46c |
| institution | Directory Open Access Journal |
| issn | 1746-1596 |
| language | English |
| last_indexed | 2024-04-12T09:53:36Z |
| publishDate | 2017-07-01 |
| publisher | BMC |
| record_format | Article |
| series | Diagnostic Pathology |
| spelling | doaj.art-ac8440886d944b0f85b02f949433d46c2022-12-22T03:37:47ZengBMCDiagnostic Pathology1746-15962017-07-011211810.1186/s13000-017-0634-3Detection of 22 common leukemic fusion genes using a single-step multiplex qRT-PCR-based assayXiaodong Lyu0Xianwei Wang1Lina Zhang2Zhenzhu Chen3Yu Zhao4Jieying Hu5Ruihua Fan6Yongping Song7School of Basic Medical Sciences, Zhengzhou UniversityCentral Laboratory, the Affiliated Cancer Hospital of Zhengzhou University; Henan Cancer HospitalDepartment of Hematology, the Affiliated Cancer Hospital of Zhengzhou University; Henan Cancer HospitalDepartment of Hematology, the Affiliated Cancer Hospital of Zhengzhou University; Henan Cancer HospitalCentral Laboratory, the Affiliated Cancer Hospital of Zhengzhou University; Henan Cancer HospitalCentral Laboratory, the Affiliated Cancer Hospital of Zhengzhou University; Henan Cancer HospitalCentral Laboratory, the Affiliated Cancer Hospital of Zhengzhou University; Henan Cancer HospitalDepartment of Hematology, the Affiliated Cancer Hospital of Zhengzhou University; Henan Cancer HospitalAbstract Background Fusion genes generated from chromosomal translocation play an important role in hematological malignancies. Detection of fusion genes currently employ use of either conventional RT-PCR methods or fluorescent in situ hybridization (FISH), where both methods involve tedious methodologies and require prior characterization of chromosomal translocation events as determined by cytogenetic analysis. In this study, we describe a real-time quantitative reverse transcription PCR (qRT-PCR)-based multi-fusion gene screening method with the capacity to detect 22 fusion genes commonly found in leukemia. This method does not require pre-characterization of gene translocation events, thereby facilitating immediate diagnosis and therapeutic management. Methods We performed fluorescent qRT-PCR (F-qRT-PCR) using a commercially-available multi-fusion gene detection kit on a patient cohort of 345 individuals comprising 108 cases diagnosed with acute myeloid leukemia (AML) for initial evaluation; remaining patients within the cohort were assayed for confirmatory diagnosis. Results obtained by F-qRT-PCR were compared alongside patient analysis by cytogenetic characterization. Results Gene translocations detected by F-qRT-PCR in AML cases were diagnosed in 69.4% of the patient cohort, which was comparatively similar to 68.5% as diagnosed by cytogenetic analysis, thereby demonstrating 99.1% concordance. Overall gene fusion was detected in 53.7% of the overall patient population by F-qRT-PCR, 52.9% by cytogenetic prediction in leukemia, and 9.1% in non-leukemia patients by both methods. The overall concordance rate was calculated to be 99.0%. Fusion genes were detected by F-qRT-PCR in 97.3% of patients with CML, followed by 69.4% with AML, 33.3% with acute lymphoblastic leukemia (ALL), 9.1% with myelodysplastic syndromes (MDS), and 0% with chronic lymphocytic leukemia (CLL). Conclusions We describe the use of a F-qRT-PCR-based multi-fusion gene screening method as an efficient one-step diagnostic procedure as an effective alternative to lengthy conventional diagnostic procedures requiring both cytogenetic analysis followed by targeted quantitative reverse transcription (qRT-PCR) methods, thus allowing timely patient management.http://link.springer.com/article/10.1186/s13000-017-0634-3LeukemiaF-qRT-PCRAMLMDSCLLALL |
| spellingShingle | Xiaodong Lyu Xianwei Wang Lina Zhang Zhenzhu Chen Yu Zhao Jieying Hu Ruihua Fan Yongping Song Detection of 22 common leukemic fusion genes using a single-step multiplex qRT-PCR-based assay Diagnostic Pathology Leukemia F-qRT-PCR AML MDS CLL ALL |
| title | Detection of 22 common leukemic fusion genes using a single-step multiplex qRT-PCR-based assay |
| title_full | Detection of 22 common leukemic fusion genes using a single-step multiplex qRT-PCR-based assay |
| title_fullStr | Detection of 22 common leukemic fusion genes using a single-step multiplex qRT-PCR-based assay |
| title_full_unstemmed | Detection of 22 common leukemic fusion genes using a single-step multiplex qRT-PCR-based assay |
| title_short | Detection of 22 common leukemic fusion genes using a single-step multiplex qRT-PCR-based assay |
| title_sort | detection of 22 common leukemic fusion genes using a single step multiplex qrt pcr based assay |
| topic | Leukemia F-qRT-PCR AML MDS CLL ALL |
| url | http://link.springer.com/article/10.1186/s13000-017-0634-3 |
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