Algorithm Optimization in Methylation Detection with Multiple RT-qPCR

Lele Song, Yuemin Li, Jia Jia, Guangpeng Zhou, Jianming Wang, Qian Kang, Peng Jin, Jianqiu Sheng, Guoxiang Cai, Sanjun Cai and Xiaoliang Han

PLOS ONE, 2016, vol. 11, issue 11, 1-14

Abstract: Epigenetic markers based on differential methylation of DNA sequences are used in cancer screening and diagnostics. Detection of abnormal methylation at specific loci by real-time quantitative polymerase chain reaction (RT-qPCR) has been developed to enable high-throughput cancer screening. For tests that combine the results of multiple PCR replicates into a single reportable result, both individual PCR cutoff and weighting of the individual PCR result are essential to test outcome. In this opportunistic screening study, we tested samples from 1133 patients using the triplicate Epi proColon assay with various algorithms and compared it with the newly developed single replicate SensiColon assay that measures methylation status of the same SEPT9 gene sequence. The Epi proColon test approved by the US FDA (1/3 algorithm) showed the highest sensitivity (82.4%) at a lower specificity (82.0%) compared with the Epi proColon 2.0 CE version with 2/3 algorithm (75.1% sensitivity, 97.1% specificity) or 1/1 algorithm (71.3% sensitivity, 92.7% specificity). No significant difference in performance was found between the Epi proColon 2.0 CE and the SensiColon assays. The choice of algorithm must depend on specific test usage, including screening and early detection. These considerations allow one to choose the optimal algorithm to maximize the test performance. We hope this study can help to optimize the methylation detection in cancer screening and early detection.

Date: 2016
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Persistent link: https://EconPapers.repec.org/RePEc:plo:pone00:0163333

DOI: 10.1371/journal.pone.0163333

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