Development of NIST standard reference material 2373: Genomic DNA standards for HER2 measurements

Q1 Biochemistry, Genetics and Molecular Biology

Biomolecular Detection and Quantification Pub Date : 2016-06-01 DOI:10.1016/j.bdq.2016.02.001

Hua-Jun He , Jamie L. Almeida , Steve P. Lund , Carolyn R. Steffen , Steve Choquette , Kenneth D. Cole

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引用次数: 19

Abstract

NIST standard reference material (SRM) 2373 was developed to improve the measurements of the HER2 gene amplification in DNA samples. SRM 2373 consists of genomic DNA extracted from five breast cancer cell lines with different amounts of amplification of the HER2 gene. The five components are derived from the human cell lines SK-BR-3, MDA-MB-231, MDA-MB-361, MDA-MB-453, and BT-474. The certified values are the ratios of the HER2 gene copy numbers to the copy numbers of selected reference genes DCK, EIF5B, RPS27A, and PMM1. The ratios were measured using quantitative polymerase chain reaction and digital PCR, methods that gave similar ratios. The five components of SRM 2373 have certified HER2 amplification ratios that range from 1.3 to 17.7. The stability and homogeneity of the reference materials were shown by repeated measurements over a period of several years. SRM 2373 is a well characterized genomic DNA reference material that can be used to improve the confidence of the measurements of HER2 gene copy number.

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开发NIST标准参考物质2373:用于HER2测量的基因组DNA标准

开发了NIST标准参比物质(SRM) 2373，以改进DNA样品中HER2基因扩增的测量。SRM 2373由从5种不同HER2基因扩增量的乳腺癌细胞系中提取的基因组DNA组成。这五种成分分别来源于人细胞系SK-BR-3、MDA-MB-231、MDA-MB-361、MDA-MB-453和BT-474。认证值为HER2基因拷贝数与选定内参基因DCK、EIF5B、RPS27A、PMM1拷贝数之比。使用定量聚合酶链反应和数字PCR方法测量比率，方法给出相似的比率。SRM 2373的五种组分具有经认证的HER2扩增比，范围从1.3到17.7。通过数年的反复测量，证明了标准物质的稳定性和均匀性。SRM 2373是一种表征良好的基因组DNA参考物质，可用于提高HER2基因拷贝数测量的置信度。

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