Development of a highly sensitive method for detection of FLT3D835Y

Background Acute myeloid leukemia (AML) is a malignant hematological neoplasm of myeloid progenitor cells. Mutations of FLT3 in its tyrosine kinase domain (FLT3-TKD) are found in ~ 8% of patients with AML, with D835Y as the most common substitution. This mutation activates survival signals that driv...
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Autor*in:	Guo, Yao [verfasserIn] Sun, Honghua [verfasserIn] Zhang, Dengyang [verfasserIn] Zhao, Yuming [verfasserIn] Shi, Mingxia [verfasserIn] Yang, Ming [verfasserIn] Xing, Shu [verfasserIn] Fu, Xueqi [verfasserIn] Bin, Ting [verfasserIn] Lu, Bo [verfasserIn] Wu, Shunjie [verfasserIn] Xu, Xiaojun [verfasserIn] Xu, Xuesong [verfasserIn] Chen, Yun [verfasserIn] Zhao, Zhizhuang Joe [verfasserIn]

Format:	E-Artikel
Sprache:	Englisch

Erschienen:	2020

Schlagwörter:	Tyrosine kinase Acute myeloid leukemia FLT3-TKD Detection of mutations

Übergeordnetes Werk:	Enthalten in: Biomarker Research - London : Biomed Central, 2013, 8(2020), 1 vom: 12. Aug.
Übergeordnetes Werk:	volume:8 ; year:2020 ; number:1 ; day:12 ; month:08

Links:	Volltext

DOI / URN:	10.1186/s40364-020-00210-7

Katalog-ID:	SPR040635910

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520			\|a Background Acute myeloid leukemia (AML) is a malignant hematological neoplasm of myeloid progenitor cells. Mutations of FLT3 in its tyrosine kinase domain (FLT3-TKD) are found in ~ 8% of patients with AML, with D835Y as the most common substitution. This mutation activates survival signals that drives the disease and is resistant to the first generation FLT3 inhibitors. Development of a highly sensitive method to detect FLT3D835Y is important to direct therapeutic options, predict prognosis, and monitor minimal residual disease in patients with AML. Methods and results In the present study, we developed a highly sensitive FLT3D835Y detection method by using the restriction fragment nested allele-specific PCR technique. The method consists of three steps: 1) initial amplification of DNA samples with PCR primers surrounding the FLT3D835Y mutation site, 2) digestion of the PCR products with restriction enzyme EcoRV that only cleaves the wild type allele, and 3) detection of FLT3D835Y by allele-specific PCR with nested primers. We were able to detect FLT3D835Y with a sensitivity of 0.001% by using purified plasmid DNAs and blood cell DNAs containing known proportions of FLT3D835Y. We analyzed blood cell DNA samples from 64 patients with AML and found six FLT3D835Y-positive cases, two of which could not be detected by conventional DNA sequencing methods. Importantly, the method was able to detect FLT3D835Y in a sample collected from a relapsed patient while the patient was in complete remission with negative MRD determined by flow cytometry. Therefore, our RFN-AS-PCR detected MRD after treatment that was missed by flow cytometry and Sanger DNA sequencing, by conventional methods. Conclusions We have developed a simple and highly sensitive method that will allow for detection of FLT3D835Y at a very low level. This method may have major clinical implications for treatment of AML.
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700	1		\|a Zhao, Zhizhuang Joe \|e verfasserin \|4 aut
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allfields	10.1186/s40364-020-00210-7 doi (DE-627)SPR040635910 (SPR)s40364-020-00210-7-e DE-627 ger DE-627 rakwb eng 570 610 ASE Guo, Yao verfasserin aut Development of a highly sensitive method for detection of FLT3D835Y 2020 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Background Acute myeloid leukemia (AML) is a malignant hematological neoplasm of myeloid progenitor cells. Mutations of FLT3 in its tyrosine kinase domain (FLT3-TKD) are found in ~ 8% of patients with AML, with D835Y as the most common substitution. This mutation activates survival signals that drives the disease and is resistant to the first generation FLT3 inhibitors. Development of a highly sensitive method to detect FLT3D835Y is important to direct therapeutic options, predict prognosis, and monitor minimal residual disease in patients with AML. Methods and results In the present study, we developed a highly sensitive FLT3D835Y detection method by using the restriction fragment nested allele-specific PCR technique. The method consists of three steps: 1) initial amplification of DNA samples with PCR primers surrounding the FLT3D835Y mutation site, 2) digestion of the PCR products with restriction enzyme EcoRV that only cleaves the wild type allele, and 3) detection of FLT3D835Y by allele-specific PCR with nested primers. We were able to detect FLT3D835Y with a sensitivity of 0.001% by using purified plasmid DNAs and blood cell DNAs containing known proportions of FLT3D835Y. We analyzed blood cell DNA samples from 64 patients with AML and found six FLT3D835Y-positive cases, two of which could not be detected by conventional DNA sequencing methods. Importantly, the method was able to detect FLT3D835Y in a sample collected from a relapsed patient while the patient was in complete remission with negative MRD determined by flow cytometry. Therefore, our RFN-AS-PCR detected MRD after treatment that was missed by flow cytometry and Sanger DNA sequencing, by conventional methods. Conclusions We have developed a simple and highly sensitive method that will allow for detection of FLT3D835Y at a very low level. This method may have major clinical implications for treatment of AML. Tyrosine kinase (dpeaa)DE-He213 Acute myeloid leukemia (dpeaa)DE-He213 FLT3-TKD (dpeaa)DE-He213 Detection of mutations (dpeaa)DE-He213 Sun, Honghua verfasserin aut Zhang, Dengyang verfasserin aut Zhao, Yuming verfasserin aut Shi, Mingxia verfasserin aut Yang, Ming verfasserin aut Xing, Shu verfasserin aut Fu, Xueqi verfasserin aut Bin, Ting verfasserin aut Lu, Bo verfasserin aut Wu, Shunjie verfasserin aut Xu, Xiaojun verfasserin aut Xu, Xuesong verfasserin aut Chen, Yun verfasserin aut Zhao, Zhizhuang Joe verfasserin aut Enthalten in Biomarker Research London : Biomed Central, 2013 8(2020), 1 vom: 12. Aug. (DE-627)735133530 (DE-600)2699926-2 2050-7771 nnns volume:8 year:2020 number:1 day:12 month:08 https://dx.doi.org/10.1186/s40364-020-00210-7 kostenfrei Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 8 2020 1 12 08
spelling	10.1186/s40364-020-00210-7 doi (DE-627)SPR040635910 (SPR)s40364-020-00210-7-e DE-627 ger DE-627 rakwb eng 570 610 ASE Guo, Yao verfasserin aut Development of a highly sensitive method for detection of FLT3D835Y 2020 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Background Acute myeloid leukemia (AML) is a malignant hematological neoplasm of myeloid progenitor cells. Mutations of FLT3 in its tyrosine kinase domain (FLT3-TKD) are found in ~ 8% of patients with AML, with D835Y as the most common substitution. This mutation activates survival signals that drives the disease and is resistant to the first generation FLT3 inhibitors. Development of a highly sensitive method to detect FLT3D835Y is important to direct therapeutic options, predict prognosis, and monitor minimal residual disease in patients with AML. Methods and results In the present study, we developed a highly sensitive FLT3D835Y detection method by using the restriction fragment nested allele-specific PCR technique. The method consists of three steps: 1) initial amplification of DNA samples with PCR primers surrounding the FLT3D835Y mutation site, 2) digestion of the PCR products with restriction enzyme EcoRV that only cleaves the wild type allele, and 3) detection of FLT3D835Y by allele-specific PCR with nested primers. We were able to detect FLT3D835Y with a sensitivity of 0.001% by using purified plasmid DNAs and blood cell DNAs containing known proportions of FLT3D835Y. We analyzed blood cell DNA samples from 64 patients with AML and found six FLT3D835Y-positive cases, two of which could not be detected by conventional DNA sequencing methods. Importantly, the method was able to detect FLT3D835Y in a sample collected from a relapsed patient while the patient was in complete remission with negative MRD determined by flow cytometry. Therefore, our RFN-AS-PCR detected MRD after treatment that was missed by flow cytometry and Sanger DNA sequencing, by conventional methods. Conclusions We have developed a simple and highly sensitive method that will allow for detection of FLT3D835Y at a very low level. This method may have major clinical implications for treatment of AML. Tyrosine kinase (dpeaa)DE-He213 Acute myeloid leukemia (dpeaa)DE-He213 FLT3-TKD (dpeaa)DE-He213 Detection of mutations (dpeaa)DE-He213 Sun, Honghua verfasserin aut Zhang, Dengyang verfasserin aut Zhao, Yuming verfasserin aut Shi, Mingxia verfasserin aut Yang, Ming verfasserin aut Xing, Shu verfasserin aut Fu, Xueqi verfasserin aut Bin, Ting verfasserin aut Lu, Bo verfasserin aut Wu, Shunjie verfasserin aut Xu, Xiaojun verfasserin aut Xu, Xuesong verfasserin aut Chen, Yun verfasserin aut Zhao, Zhizhuang Joe verfasserin aut Enthalten in Biomarker Research London : Biomed Central, 2013 8(2020), 1 vom: 12. Aug. (DE-627)735133530 (DE-600)2699926-2 2050-7771 nnns volume:8 year:2020 number:1 day:12 month:08 https://dx.doi.org/10.1186/s40364-020-00210-7 kostenfrei Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 8 2020 1 12 08
allfields_unstemmed	10.1186/s40364-020-00210-7 doi (DE-627)SPR040635910 (SPR)s40364-020-00210-7-e DE-627 ger DE-627 rakwb eng 570 610 ASE Guo, Yao verfasserin aut Development of a highly sensitive method for detection of FLT3D835Y 2020 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Background Acute myeloid leukemia (AML) is a malignant hematological neoplasm of myeloid progenitor cells. Mutations of FLT3 in its tyrosine kinase domain (FLT3-TKD) are found in ~ 8% of patients with AML, with D835Y as the most common substitution. This mutation activates survival signals that drives the disease and is resistant to the first generation FLT3 inhibitors. Development of a highly sensitive method to detect FLT3D835Y is important to direct therapeutic options, predict prognosis, and monitor minimal residual disease in patients with AML. Methods and results In the present study, we developed a highly sensitive FLT3D835Y detection method by using the restriction fragment nested allele-specific PCR technique. The method consists of three steps: 1) initial amplification of DNA samples with PCR primers surrounding the FLT3D835Y mutation site, 2) digestion of the PCR products with restriction enzyme EcoRV that only cleaves the wild type allele, and 3) detection of FLT3D835Y by allele-specific PCR with nested primers. We were able to detect FLT3D835Y with a sensitivity of 0.001% by using purified plasmid DNAs and blood cell DNAs containing known proportions of FLT3D835Y. We analyzed blood cell DNA samples from 64 patients with AML and found six FLT3D835Y-positive cases, two of which could not be detected by conventional DNA sequencing methods. Importantly, the method was able to detect FLT3D835Y in a sample collected from a relapsed patient while the patient was in complete remission with negative MRD determined by flow cytometry. Therefore, our RFN-AS-PCR detected MRD after treatment that was missed by flow cytometry and Sanger DNA sequencing, by conventional methods. Conclusions We have developed a simple and highly sensitive method that will allow for detection of FLT3D835Y at a very low level. This method may have major clinical implications for treatment of AML. Tyrosine kinase (dpeaa)DE-He213 Acute myeloid leukemia (dpeaa)DE-He213 FLT3-TKD (dpeaa)DE-He213 Detection of mutations (dpeaa)DE-He213 Sun, Honghua verfasserin aut Zhang, Dengyang verfasserin aut Zhao, Yuming verfasserin aut Shi, Mingxia verfasserin aut Yang, Ming verfasserin aut Xing, Shu verfasserin aut Fu, Xueqi verfasserin aut Bin, Ting verfasserin aut Lu, Bo verfasserin aut Wu, Shunjie verfasserin aut Xu, Xiaojun verfasserin aut Xu, Xuesong verfasserin aut Chen, Yun verfasserin aut Zhao, Zhizhuang Joe verfasserin aut Enthalten in Biomarker Research London : Biomed Central, 2013 8(2020), 1 vom: 12. Aug. (DE-627)735133530 (DE-600)2699926-2 2050-7771 nnns volume:8 year:2020 number:1 day:12 month:08 https://dx.doi.org/10.1186/s40364-020-00210-7 kostenfrei Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 8 2020 1 12 08
allfieldsGer	10.1186/s40364-020-00210-7 doi (DE-627)SPR040635910 (SPR)s40364-020-00210-7-e DE-627 ger DE-627 rakwb eng 570 610 ASE Guo, Yao verfasserin aut Development of a highly sensitive method for detection of FLT3D835Y 2020 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Background Acute myeloid leukemia (AML) is a malignant hematological neoplasm of myeloid progenitor cells. Mutations of FLT3 in its tyrosine kinase domain (FLT3-TKD) are found in ~ 8% of patients with AML, with D835Y as the most common substitution. This mutation activates survival signals that drives the disease and is resistant to the first generation FLT3 inhibitors. Development of a highly sensitive method to detect FLT3D835Y is important to direct therapeutic options, predict prognosis, and monitor minimal residual disease in patients with AML. Methods and results In the present study, we developed a highly sensitive FLT3D835Y detection method by using the restriction fragment nested allele-specific PCR technique. The method consists of three steps: 1) initial amplification of DNA samples with PCR primers surrounding the FLT3D835Y mutation site, 2) digestion of the PCR products with restriction enzyme EcoRV that only cleaves the wild type allele, and 3) detection of FLT3D835Y by allele-specific PCR with nested primers. We were able to detect FLT3D835Y with a sensitivity of 0.001% by using purified plasmid DNAs and blood cell DNAs containing known proportions of FLT3D835Y. We analyzed blood cell DNA samples from 64 patients with AML and found six FLT3D835Y-positive cases, two of which could not be detected by conventional DNA sequencing methods. Importantly, the method was able to detect FLT3D835Y in a sample collected from a relapsed patient while the patient was in complete remission with negative MRD determined by flow cytometry. Therefore, our RFN-AS-PCR detected MRD after treatment that was missed by flow cytometry and Sanger DNA sequencing, by conventional methods. Conclusions We have developed a simple and highly sensitive method that will allow for detection of FLT3D835Y at a very low level. This method may have major clinical implications for treatment of AML. Tyrosine kinase (dpeaa)DE-He213 Acute myeloid leukemia (dpeaa)DE-He213 FLT3-TKD (dpeaa)DE-He213 Detection of mutations (dpeaa)DE-He213 Sun, Honghua verfasserin aut Zhang, Dengyang verfasserin aut Zhao, Yuming verfasserin aut Shi, Mingxia verfasserin aut Yang, Ming verfasserin aut Xing, Shu verfasserin aut Fu, Xueqi verfasserin aut Bin, Ting verfasserin aut Lu, Bo verfasserin aut Wu, Shunjie verfasserin aut Xu, Xiaojun verfasserin aut Xu, Xuesong verfasserin aut Chen, Yun verfasserin aut Zhao, Zhizhuang Joe verfasserin aut Enthalten in Biomarker Research London : Biomed Central, 2013 8(2020), 1 vom: 12. Aug. (DE-627)735133530 (DE-600)2699926-2 2050-7771 nnns volume:8 year:2020 number:1 day:12 month:08 https://dx.doi.org/10.1186/s40364-020-00210-7 kostenfrei Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 8 2020 1 12 08
allfieldsSound	10.1186/s40364-020-00210-7 doi (DE-627)SPR040635910 (SPR)s40364-020-00210-7-e DE-627 ger DE-627 rakwb eng 570 610 ASE Guo, Yao verfasserin aut Development of a highly sensitive method for detection of FLT3D835Y 2020 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Background Acute myeloid leukemia (AML) is a malignant hematological neoplasm of myeloid progenitor cells. Mutations of FLT3 in its tyrosine kinase domain (FLT3-TKD) are found in ~ 8% of patients with AML, with D835Y as the most common substitution. This mutation activates survival signals that drives the disease and is resistant to the first generation FLT3 inhibitors. Development of a highly sensitive method to detect FLT3D835Y is important to direct therapeutic options, predict prognosis, and monitor minimal residual disease in patients with AML. Methods and results In the present study, we developed a highly sensitive FLT3D835Y detection method by using the restriction fragment nested allele-specific PCR technique. The method consists of three steps: 1) initial amplification of DNA samples with PCR primers surrounding the FLT3D835Y mutation site, 2) digestion of the PCR products with restriction enzyme EcoRV that only cleaves the wild type allele, and 3) detection of FLT3D835Y by allele-specific PCR with nested primers. We were able to detect FLT3D835Y with a sensitivity of 0.001% by using purified plasmid DNAs and blood cell DNAs containing known proportions of FLT3D835Y. We analyzed blood cell DNA samples from 64 patients with AML and found six FLT3D835Y-positive cases, two of which could not be detected by conventional DNA sequencing methods. Importantly, the method was able to detect FLT3D835Y in a sample collected from a relapsed patient while the patient was in complete remission with negative MRD determined by flow cytometry. Therefore, our RFN-AS-PCR detected MRD after treatment that was missed by flow cytometry and Sanger DNA sequencing, by conventional methods. Conclusions We have developed a simple and highly sensitive method that will allow for detection of FLT3D835Y at a very low level. This method may have major clinical implications for treatment of AML. Tyrosine kinase (dpeaa)DE-He213 Acute myeloid leukemia (dpeaa)DE-He213 FLT3-TKD (dpeaa)DE-He213 Detection of mutations (dpeaa)DE-He213 Sun, Honghua verfasserin aut Zhang, Dengyang verfasserin aut Zhao, Yuming verfasserin aut Shi, Mingxia verfasserin aut Yang, Ming verfasserin aut Xing, Shu verfasserin aut Fu, Xueqi verfasserin aut Bin, Ting verfasserin aut Lu, Bo verfasserin aut Wu, Shunjie verfasserin aut Xu, Xiaojun verfasserin aut Xu, Xuesong verfasserin aut Chen, Yun verfasserin aut Zhao, Zhizhuang Joe verfasserin aut Enthalten in Biomarker Research London : Biomed Central, 2013 8(2020), 1 vom: 12. Aug. (DE-627)735133530 (DE-600)2699926-2 2050-7771 nnns volume:8 year:2020 number:1 day:12 month:08 https://dx.doi.org/10.1186/s40364-020-00210-7 kostenfrei Volltext GBV_USEFLAG_A SYSFLAG_A GBV_SPRINGER SSG-OLC-PHA GBV_ILN_11 GBV_ILN_20 GBV_ILN_22 GBV_ILN_23 GBV_ILN_24 GBV_ILN_39 GBV_ILN_40 GBV_ILN_60 GBV_ILN_62 GBV_ILN_63 GBV_ILN_65 GBV_ILN_69 GBV_ILN_70 GBV_ILN_73 GBV_ILN_74 GBV_ILN_95 GBV_ILN_105 GBV_ILN_110 GBV_ILN_151 GBV_ILN_161 GBV_ILN_170 GBV_ILN_206 GBV_ILN_213 GBV_ILN_230 GBV_ILN_285 GBV_ILN_293 GBV_ILN_602 GBV_ILN_2003 GBV_ILN_2014 GBV_ILN_4012 GBV_ILN_4037 GBV_ILN_4112 GBV_ILN_4125 GBV_ILN_4126 GBV_ILN_4249 GBV_ILN_4305 GBV_ILN_4306 GBV_ILN_4307 GBV_ILN_4313 GBV_ILN_4322 GBV_ILN_4323 GBV_ILN_4324 GBV_ILN_4325 GBV_ILN_4338 GBV_ILN_4367 GBV_ILN_4700 AR 8 2020 1 12 08
language	English
source	Enthalten in Biomarker Research 8(2020), 1 vom: 12. Aug. volume:8 year:2020 number:1 day:12 month:08
sourceStr	Enthalten in Biomarker Research 8(2020), 1 vom: 12. Aug. volume:8 year:2020 number:1 day:12 month:08
format_phy_str_mv	Article
institution	findex.gbv.de
topic_facet	Tyrosine kinase Acute myeloid leukemia FLT3-TKD Detection of mutations
dewey-raw	570
isfreeaccess_bool	true
container_title	Biomarker Research
authorswithroles_txt_mv	Guo, Yao @@aut@@ Sun, Honghua @@aut@@ Zhang, Dengyang @@aut@@ Zhao, Yuming @@aut@@ Shi, Mingxia @@aut@@ Yang, Ming @@aut@@ Xing, Shu @@aut@@ Fu, Xueqi @@aut@@ Bin, Ting @@aut@@ Lu, Bo @@aut@@ Wu, Shunjie @@aut@@ Xu, Xiaojun @@aut@@ Xu, Xuesong @@aut@@ Chen, Yun @@aut@@ Zhao, Zhizhuang Joe @@aut@@
publishDateDaySort_date	2020-08-12T00:00:00Z
hierarchy_top_id	735133530
dewey-sort	3570
id	SPR040635910
language_de	englisch
fullrecord	<?xml version="1.0" encoding="UTF-8"?><collection xmlns="http://www.loc.gov/MARC21/slim"><record><leader>01000caa a22002652 4500</leader><controlfield tag="001">SPR040635910</controlfield><controlfield tag="003">DE-627</controlfield><controlfield tag="005">20230519174957.0</controlfield><controlfield tag="007">cr uuu---uuuuu</controlfield><controlfield tag="008">201007s2020 xx \|\|\|\|\|o 00\| \|\|eng c</controlfield><datafield tag="024" ind1="7" ind2=" "><subfield code="a">10.1186/s40364-020-00210-7</subfield><subfield code="2">doi</subfield></datafield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(DE-627)SPR040635910</subfield></datafield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(SPR)s40364-020-00210-7-e</subfield></datafield><datafield tag="040" ind1=" " ind2=" "><subfield code="a">DE-627</subfield><subfield code="b">ger</subfield><subfield code="c">DE-627</subfield><subfield code="e">rakwb</subfield></datafield><datafield tag="041" ind1=" " ind2=" "><subfield code="a">eng</subfield></datafield><datafield tag="082" ind1="0" ind2="4"><subfield code="a">570</subfield><subfield code="a">610</subfield><subfield code="q">ASE</subfield></datafield><datafield tag="100" ind1="1" ind2=" "><subfield code="a">Guo, Yao</subfield><subfield code="e">verfasserin</subfield><subfield code="4">aut</subfield></datafield><datafield tag="245" ind1="1" ind2="0"><subfield code="a">Development of a highly sensitive method for detection of FLT3D835Y</subfield></datafield><datafield tag="264" ind1=" " ind2="1"><subfield code="c">2020</subfield></datafield><datafield tag="336" ind1=" " ind2=" "><subfield code="a">Text</subfield><subfield code="b">txt</subfield><subfield code="2">rdacontent</subfield></datafield><datafield tag="337" ind1=" " ind2=" "><subfield code="a">Computermedien</subfield><subfield code="b">c</subfield><subfield code="2">rdamedia</subfield></datafield><datafield tag="338" ind1=" " ind2=" "><subfield code="a">Online-Ressource</subfield><subfield code="b">cr</subfield><subfield code="2">rdacarrier</subfield></datafield><datafield tag="520" ind1=" " ind2=" "><subfield code="a">Background Acute myeloid leukemia (AML) is a malignant hematological neoplasm of myeloid progenitor cells. Mutations of FLT3 in its tyrosine kinase domain (FLT3-TKD) are found in ~ 8% of patients with AML, with D835Y as the most common substitution. This mutation activates survival signals that drives the disease and is resistant to the first generation FLT3 inhibitors. Development of a highly sensitive method to detect FLT3D835Y is important to direct therapeutic options, predict prognosis, and monitor minimal residual disease in patients with AML. Methods and results In the present study, we developed a highly sensitive FLT3D835Y detection method by using the restriction fragment nested allele-specific PCR technique. The method consists of three steps: 1) initial amplification of DNA samples with PCR primers surrounding the FLT3D835Y mutation site, 2) digestion of the PCR products with restriction enzyme EcoRV that only cleaves the wild type allele, and 3) detection of FLT3D835Y by allele-specific PCR with nested primers. We were able to detect FLT3D835Y with a sensitivity of 0.001% by using purified plasmid DNAs and blood cell DNAs containing known proportions of FLT3D835Y. We analyzed blood cell DNA samples from 64 patients with AML and found six FLT3D835Y-positive cases, two of which could not be detected by conventional DNA sequencing methods. Importantly, the method was able to detect FLT3D835Y in a sample collected from a relapsed patient while the patient was in complete remission with negative MRD determined by flow cytometry. Therefore, our RFN-AS-PCR detected MRD after treatment that was missed by flow cytometry and Sanger DNA sequencing, by conventional methods. Conclusions We have developed a simple and highly sensitive method that will allow for detection of FLT3D835Y at a very low level. 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author	Guo, Yao
spellingShingle	Guo, Yao ddc 570 misc Tyrosine kinase misc Acute myeloid leukemia misc FLT3-TKD misc Detection of mutations Development of a highly sensitive method for detection of FLT3D835Y
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topic_title	570 610 ASE Development of a highly sensitive method for detection of FLT3D835Y Tyrosine kinase (dpeaa)DE-He213 Acute myeloid leukemia (dpeaa)DE-He213 FLT3-TKD (dpeaa)DE-He213 Detection of mutations (dpeaa)DE-He213
topic	ddc 570 misc Tyrosine kinase misc Acute myeloid leukemia misc FLT3-TKD misc Detection of mutations
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title	Development of a highly sensitive method for detection of FLT3D835Y
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title_full	Development of a highly sensitive method for detection of FLT3D835Y
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author_browse	Guo, Yao Sun, Honghua Zhang, Dengyang Zhao, Yuming Shi, Mingxia Yang, Ming Xing, Shu Fu, Xueqi Bin, Ting Lu, Bo Wu, Shunjie Xu, Xiaojun Xu, Xuesong Chen, Yun Zhao, Zhizhuang Joe
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title_sort	development of a highly sensitive method for detection of flt3d835y
title_auth	Development of a highly sensitive method for detection of FLT3D835Y
abstract	Background Acute myeloid leukemia (AML) is a malignant hematological neoplasm of myeloid progenitor cells. Mutations of FLT3 in its tyrosine kinase domain (FLT3-TKD) are found in ~ 8% of patients with AML, with D835Y as the most common substitution. This mutation activates survival signals that drives the disease and is resistant to the first generation FLT3 inhibitors. Development of a highly sensitive method to detect FLT3D835Y is important to direct therapeutic options, predict prognosis, and monitor minimal residual disease in patients with AML. Methods and results In the present study, we developed a highly sensitive FLT3D835Y detection method by using the restriction fragment nested allele-specific PCR technique. The method consists of three steps: 1) initial amplification of DNA samples with PCR primers surrounding the FLT3D835Y mutation site, 2) digestion of the PCR products with restriction enzyme EcoRV that only cleaves the wild type allele, and 3) detection of FLT3D835Y by allele-specific PCR with nested primers. We were able to detect FLT3D835Y with a sensitivity of 0.001% by using purified plasmid DNAs and blood cell DNAs containing known proportions of FLT3D835Y. We analyzed blood cell DNA samples from 64 patients with AML and found six FLT3D835Y-positive cases, two of which could not be detected by conventional DNA sequencing methods. Importantly, the method was able to detect FLT3D835Y in a sample collected from a relapsed patient while the patient was in complete remission with negative MRD determined by flow cytometry. Therefore, our RFN-AS-PCR detected MRD after treatment that was missed by flow cytometry and Sanger DNA sequencing, by conventional methods. Conclusions We have developed a simple and highly sensitive method that will allow for detection of FLT3D835Y at a very low level. This method may have major clinical implications for treatment of AML.
abstractGer	Background Acute myeloid leukemia (AML) is a malignant hematological neoplasm of myeloid progenitor cells. Mutations of FLT3 in its tyrosine kinase domain (FLT3-TKD) are found in ~ 8% of patients with AML, with D835Y as the most common substitution. This mutation activates survival signals that drives the disease and is resistant to the first generation FLT3 inhibitors. Development of a highly sensitive method to detect FLT3D835Y is important to direct therapeutic options, predict prognosis, and monitor minimal residual disease in patients with AML. Methods and results In the present study, we developed a highly sensitive FLT3D835Y detection method by using the restriction fragment nested allele-specific PCR technique. The method consists of three steps: 1) initial amplification of DNA samples with PCR primers surrounding the FLT3D835Y mutation site, 2) digestion of the PCR products with restriction enzyme EcoRV that only cleaves the wild type allele, and 3) detection of FLT3D835Y by allele-specific PCR with nested primers. We were able to detect FLT3D835Y with a sensitivity of 0.001% by using purified plasmid DNAs and blood cell DNAs containing known proportions of FLT3D835Y. We analyzed blood cell DNA samples from 64 patients with AML and found six FLT3D835Y-positive cases, two of which could not be detected by conventional DNA sequencing methods. Importantly, the method was able to detect FLT3D835Y in a sample collected from a relapsed patient while the patient was in complete remission with negative MRD determined by flow cytometry. Therefore, our RFN-AS-PCR detected MRD after treatment that was missed by flow cytometry and Sanger DNA sequencing, by conventional methods. Conclusions We have developed a simple and highly sensitive method that will allow for detection of FLT3D835Y at a very low level. This method may have major clinical implications for treatment of AML.
abstract_unstemmed	Background Acute myeloid leukemia (AML) is a malignant hematological neoplasm of myeloid progenitor cells. Mutations of FLT3 in its tyrosine kinase domain (FLT3-TKD) are found in ~ 8% of patients with AML, with D835Y as the most common substitution. This mutation activates survival signals that drives the disease and is resistant to the first generation FLT3 inhibitors. Development of a highly sensitive method to detect FLT3D835Y is important to direct therapeutic options, predict prognosis, and monitor minimal residual disease in patients with AML. Methods and results In the present study, we developed a highly sensitive FLT3D835Y detection method by using the restriction fragment nested allele-specific PCR technique. The method consists of three steps: 1) initial amplification of DNA samples with PCR primers surrounding the FLT3D835Y mutation site, 2) digestion of the PCR products with restriction enzyme EcoRV that only cleaves the wild type allele, and 3) detection of FLT3D835Y by allele-specific PCR with nested primers. We were able to detect FLT3D835Y with a sensitivity of 0.001% by using purified plasmid DNAs and blood cell DNAs containing known proportions of FLT3D835Y. We analyzed blood cell DNA samples from 64 patients with AML and found six FLT3D835Y-positive cases, two of which could not be detected by conventional DNA sequencing methods. Importantly, the method was able to detect FLT3D835Y in a sample collected from a relapsed patient while the patient was in complete remission with negative MRD determined by flow cytometry. Therefore, our RFN-AS-PCR detected MRD after treatment that was missed by flow cytometry and Sanger DNA sequencing, by conventional methods. Conclusions We have developed a simple and highly sensitive method that will allow for detection of FLT3D835Y at a very low level. This method may have major clinical implications for treatment of AML.
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title_short	Development of a highly sensitive method for detection of FLT3D835Y
url	https://dx.doi.org/10.1186/s40364-020-00210-7
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author2	Sun, Honghua Zhang, Dengyang Zhao, Yuming Shi, Mingxia Yang, Ming Xing, Shu Fu, Xueqi Bin, Ting Lu, Bo Wu, Shunjie Xu, Xiaojun Xu, Xuesong Chen, Yun Zhao, Zhizhuang Joe
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We were able to detect FLT3D835Y with a sensitivity of 0.001% by using purified plasmid DNAs and blood cell DNAs containing known proportions of FLT3D835Y. We analyzed blood cell DNA samples from 64 patients with AML and found six FLT3D835Y-positive cases, two of which could not be detected by conventional DNA sequencing methods. Importantly, the method was able to detect FLT3D835Y in a sample collected from a relapsed patient while the patient was in complete remission with negative MRD determined by flow cytometry. Therefore, our RFN-AS-PCR detected MRD after treatment that was missed by flow cytometry and Sanger DNA sequencing, by conventional methods. Conclusions We have developed a simple and highly sensitive method that will allow for detection of FLT3D835Y at a very low level. 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score	7.4001493

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Development of a highly sensitive method for detection of FLT3D835Y

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Zugang & Verfügbarkeit

Vorhandene Bände

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