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An Extraction-Free Method for Rapid Detection of CYP2C19 * 2/3/17 Polymorphisms in One Tube using Melting Curve Analysis
  • +8
  • Jianguang Guo,
  • Kangfeng Lin,
  • Weixin You,
  • Qinghan Li,
  • Xiangju Guo,
  • Shuai Wang,
  • Ya Bian,
  • Wenjing Ren,
  • Rui Zhang,
  • Yanping Wang,
  • Bo-An Li
Jianguang Guo
Xiamen University School of Life Sciences
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Kangfeng Lin
Xiamen University School of Life Sciences
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Weixin You
Xiamen University School of Life Sciences
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Qinghan Li
Xiamen University School of Life Sciences
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Xiangju Guo
Xiamen University School of Life Sciences
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Shuai Wang
Xiamen University School of Life Sciences
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Ya Bian
Xiamen University School of Life Sciences
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Wenjing Ren
Xiamen University School of Life Sciences
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Rui Zhang
The First Affiliated Hospital of Xiamen University
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Yanping Wang
Hubei Provincial Hospital of Traditional Chinese Medicine
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Bo-An Li
Xiamen University School of Life Sciences

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Abstract

Drug-metabolizing enzymes play an important role in the metabolism of drugs in vivo. Their activity is an important factor affecting the rate of drug metabolism, which directly determines the intensity and persistence of drug action. Patients taking medication can be divided into different metabolic types through detection of CYP2C19 drug-metabolizing enzyme gene polymorphisms, which can then be used for medication guidance for clopidogrel. Here, we describe a detection method based on real-time polymerase chain reaction. This method uses multicolor melting curve analysis to accurately identify different mutation sites and genotypes of CYP2C19 * 2, CYP2C19 * 3, and CYP2C19 * 17. The detection limit of plasmid samples was 1 copies/µl; that of genomic samples was 0.1 ng/µl. The system can detect nine types of CYP2C19 * 2/3/17 at three sites in one tube, quickly achieving detection within 1 h. Combined with the sample release agent, sample extraction was completed in 5 s, achieving rapid diagnosis without extraction for timely diagnosis and treatment. Furthermore, the system is not limited to blood samples and can also be applied to oropharyngeal and saliva samples, increasing sampling diversity and convenience. When using clinical blood samples (n=93), the detection system we established was able to quickly and accurately identify different genotypes, and the accuracy and effectiveness of the detection were confirmed by Sanger sequencing.
08 May 2023Submitted to Biotechnology Journal
09 May 2023Submission Checks Completed
09 May 2023Assigned to Editor
10 May 2023Reviewer(s) Assigned
24 May 2023Review(s) Completed, Editorial Evaluation Pending
25 May 2023Editorial Decision: Revise Major
11 Jul 20231st Revision Received
12 Jul 2023Submission Checks Completed
12 Jul 2023Assigned to Editor
12 Jul 2023Reviewer(s) Assigned
30 Jul 2023Review(s) Completed, Editorial Evaluation Pending
02 Aug 2023Editorial Decision: Accept