一种用于鉴定和定量游离DNA中EGFR T790M的超灵敏qPCR方法的开发与验证

Development and validation of an ultrasensitive qPCR method to identify and quantify EGFR T790M in cell-free DNA.

作者信息

Yuan Shenglei, Jia Nan, Lu Guofu, Lai Jinping, Liang Wenzhong, Li Lan, Zhang Chenpu, Diao Jianbo

机构信息

Bioanalytical Services Department, WuXi AppTec (Shanghai) Co. Ltd, Shanghai, China.

Bioanalytical Services Department, WuXi AppTec, Plainsboro, NJ, USA.

出版信息

Bioanalysis. 2025 Jan;17(1):49-62. doi: 10.1080/17576180.2025.2451527. Epub 2025 Jan 15.

DOI:10.1080/17576180.2025.2451527

PMID:39812332

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11749345/

Abstract

BACKGROUND

Circulating tumor DNA (ctDNA) is a promising biomarker for cancer prognosis and drug development. A major challenge in the ctDNA determination method is discriminating ctDNA from highly similar but significantly more abundant wild-type DNA sensitively and accurately.

METHOD

An ultrasensitive qPCR method termed Triple Enrichment Amplification of Mutation PCR (TEAM-PCR) was developed to detect EGFR T790M mutation.

RESULTS

EGFR T790M was quantified over the assay range of 25-10 copies/reaction in the presence of 10 wild-type copies. This method was fully validated following the essential bioanalysis guidance, with the limit of detection (LOD) being five copies/reaction.

CONCLUSION

This study established and validated a qPCR-based strategy to detect EGFR T790M mutation with ultra-high sensitivity and reliability.

摘要

背景

循环肿瘤DNA（ctDNA）是一种很有前景的癌症预后和药物开发生物标志物。ctDNA检测方法的一个主要挑战是灵敏且准确地将ctDNA与高度相似但数量多得多的野生型DNA区分开来。

方法

开发了一种超灵敏定量聚合酶链反应方法，称为突变PCR三重富集扩增（TEAM-PCR），用于检测表皮生长因子受体（EGFR）T790M突变。

结果

在存在10个野生型拷贝的情况下，EGFR T790M在25-10拷贝/反应的检测范围内被定量。该方法按照基本生物分析指南进行了全面验证，检测限为5拷贝/反应。

结论

本研究建立并验证了一种基于定量聚合酶链反应的策略，以超高灵敏度和可靠性检测EGFR T790M突变。

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一种用于鉴定和定量游离DNA中EGFR T790M的超灵敏qPCR方法的开发与验证

Development and validation of an ultrasensitive qPCR method to identify and quantify EGFR T790M in cell-free DNA.

作者信息

Yuan Shenglei, Jia Nan, Lu Guofu, Lai Jinping, Liang Wenzhong, Li Lan, Zhang Chenpu, Diao Jianbo

机构信息

Bioanalytical Services Department, WuXi AppTec (Shanghai) Co. Ltd, Shanghai, China.

Bioanalytical Services Department, WuXi AppTec, Plainsboro, NJ, USA.

出版信息

Bioanalysis. 2025 Jan;17(1):49-62. doi: 10.1080/17576180.2025.2451527. Epub 2025 Jan 15.

DOI:10.1080/17576180.2025.2451527

PMID:39812332

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11749345/

Abstract

BACKGROUND

METHOD

An ultrasensitive qPCR method termed Triple Enrichment Amplification of Mutation PCR (TEAM-PCR) was developed to detect EGFR T790M mutation.

RESULTS

CONCLUSION

This study established and validated a qPCR-based strategy to detect EGFR T790M mutation with ultra-high sensitivity and reliability.

摘要

背景

方法

开发了一种超灵敏定量聚合酶链反应方法，称为突变PCR三重富集扩增（TEAM-PCR），用于检测表皮生长因子受体（EGFR）T790M突变。

结果

在存在10个野生型拷贝的情况下，EGFR T790M在25-10拷贝/反应的检测范围内被定量。该方法按照基本生物分析指南进行了全面验证，检测限为5拷贝/反应。

结论

本研究建立并验证了一种基于定量聚合酶链反应的策略，以超高灵敏度和可靠性检测EGFR T790M突变。

一种用于鉴定和定量游离DNA中EGFR T790M的超灵敏qPCR方法的开发与验证

Development and validation of an ultrasensitive qPCR method to identify and quantify EGFR T790M in cell-free DNA.

作者信息

机构信息

出版信息

BACKGROUND

METHOD

RESULTS

CONCLUSION

背景

方法

结果

结论

相似文献

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一种用于鉴定和定量游离DNA中EGFR T790M的超灵敏qPCR方法的开发与验证

Development and validation of an ultrasensitive qPCR method to identify and quantify EGFR T790M in cell-free DNA.

作者信息

机构信息

出版信息

BACKGROUND

METHOD

RESULTS

CONCLUSION

背景

方法

结果

结论