Droplet digital PCR-based &ITEGFR &ITmutation detection with an internal quality control index to determine the quality of DNA

作者：Kim Sung Su; Choi Hyun Jeung; Kim Jin Ju; Kim M Sun; Lee In Seon; Byun Bohyun; Jia Lina; Oh Myung Ryurl; Moon Youngho; Parka Sarah; Choi Joon Seok; Chae Seoung Wan; Nam Byung Ho; Kim Jin Soo; Kim Jihun; Min Byung Soh; Lee Jae Seok; Won Jae Kyung; Cho SooYoun; Choi Yoon La; Shin Young Kee^*

来源：Scientific Reports, 2018, 8(1): 543.

DOI：10.1038/s41598-017-18642-x

In clinical translational research and molecular in vitro diagnostics, a major challenge in the detection of genetic mutations is overcoming artefactual results caused by the low-quality of formalin-fixed paraffin-embedded tissue (FFPET)-derived DNA (FFPET-DNA). Here, we propose the use of an 'internal quality control (iQC) index' as a criterion for judging the minimum quality of DNA for PCR-based analyses. In a pre-clinical study comparing the results from droplet digital PCR-based EGFR mutation test (ddEGFR test) and qPCR-based EGFR mutation test (cobas EGFR test), iQC index >= 0.5 (iQC copies >= 500, using 3.3 ng of FFPET-DNA [1,000 genome equivalents]) was established, indicating that more than half of the input DNA was amplifiable. Using this criterion, we conducted a retrospective comparative clinical study of the ddEGFR and cobas EGFR tests for the detection of EGFR mutations in non-small cell lung cancer (NSCLC) FFPET-DNA samples. Compared with the cobas EGFR test, the ddEGFR test exhibited superior analytical performance and equivalent or higher clinical performance. Furthermore, iQC index is a reliable indicator of the quality of FFPET-DNA and could be used to prevent incorrect diagnoses arising from low-quality samples.

• 出版日期2018-1-11