A novel microfluidic chip-based digital PCR method for enhanced sensitivity in the early diagnosis of colorectal cancer via mSEPT9

• Published in: Clinica chimica acta Vol. 554; p. 117781
• Main Authors: Huang, Qunfang, Xun, Zhen, Lin, Junyu, Xie, Rubing, Zhu, Chenggong, Wang, Long, Shang, Hongyan, Wu, Songhang, Ou, Qishui, Liu, Can
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01-02-2024
• Subjects: Colorectal cancer; Diagnosis; Digital PCR; Septin9; Septin9; FDA; CV; CRC; LNAs; ROC; Colorectal cancer; FOBT; Digital PCR; Diagnosis; CEA
• ISSN: 0009-8981; 1873-3492
• DOI: 10.1016/j.cca.2024.117781

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•A new approach for detecting colorectal cancer (CRC) has been developed using a microfluidic chip-based digital PCR (dPCR) method. This method focuses on detecting a specific gene called mSEPT9 and has been designed for low-concentration samples.•The chip-based dPCR method utilizes modified primers and probes for improved accuracy. In clinical testing, this method outperformed a standard qPCR kit, especially in early-stage CRC diagnosis, and its accuracy was further enhanced when combined with the carcinoembryonic antigen (CEA) test.
To enhance the sensitivity of plasma methylated Septin9 gene (mSEPT9) detection in colorectal cancer (CRC) screening, we developed a microfluidic chip-based digital PCR (dPCR) method suitable for low-concentration samples, aiming to apply it for mSEPT9 detection in CRC diagnosis.
Our microfluidic chip-based dPCR method utilized specific primers and probes with locked nucleic acids (LNAs) modifications for mSEPT9 detection. We evaluated its performance, including detection limit, specificity, and linear range, comparing it with a commercial qPCR reagent kit using the same samples (95 CRC, 23 non-CRC).
The LNAs-modified dPCR method showed a linear range of 100–104 copies/μL and a detection limit of 100 copies/μL. Clinical testing revealed that our dPCR method exhibited a sensitivity of 82.11 % and specificity of 95.65 % for CRC diagnosis, outperforming the commercial qPCR kit (sensitivity: 58.95 %, specificity: 91.30 %), particularly in Stage I with a diagnostic sensitivity of 90.91 %. Combining mSEPT9 and carcinoembryonic antigen (CEA) improved diagnostic sensitivity to 91.49 %.
Our accurate microfluidic chip-based dPCR method, especially in combination with CEA, holds promise for effective CRC screening and timely interventions, offering enhanced mSEPT9 quantification over conventional qPCR.