A highly sensitive electrochemical magneto-genosensing assay for the specific detection of a single nucleotide variation in the KRAS oncogene in human plasma
Liquid biopsy is a non-invasive and efficient technique for the detection of tumor biomarkers in biological fluids, which currently represents a new frontier in theranostics and precision medicine. Among mutations of the KRAS oncogene, p.G12D single nucleotide variation in the KRAS gene plays a cent...
| Main Authors: | , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2023-12-01
|
| Series: | Biosensors and Bioelectronics: X |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2590137023001012 |
| _version_ | 1797398475271307264 |
|---|---|
| author | Simone Fortunati Chiara Giliberti Marco Giannetto Alessandro Bertucci Sabrina Capodaglio Elena Ricciardi Patrizio Giacomini Valentina Bianchi Andrea Boni Ilaria De Munari Roberto Corradini Maria Careri |
| author_facet | Simone Fortunati Chiara Giliberti Marco Giannetto Alessandro Bertucci Sabrina Capodaglio Elena Ricciardi Patrizio Giacomini Valentina Bianchi Andrea Boni Ilaria De Munari Roberto Corradini Maria Careri |
| author_sort | Simone Fortunati |
| collection | DOAJ |
| description | Liquid biopsy is a non-invasive and efficient technique for the detection of tumor biomarkers in biological fluids, which currently represents a new frontier in theranostics and precision medicine. Among mutations of the KRAS oncogene, p.G12D single nucleotide variation in the KRAS gene plays a central role in the early diagnosis and therapeutic treatment of colorectal cancer. In this context, we developed a highly sensitive magneto-genosensing assay based on PNA capture probes immobilized on magnetic microbeads. To detect the mutation of the KRAS oncogene, two capture probe sequences recognizing wild-type and p.G12D tumor DNA were used in association with a DNA signaling probe allowing for electrochemical detection using an enzyme conjugate. The assay conditions were optimized using a 32 full-factorial design, obtaining an outstanding specificity, evidenced by a remarkably lower (>95%) signal of the singly-mismatched- compared to that of fully-complementary-DNA. Ultra-high sensitivity was achieved in 10-fold diluted human plasma reaching detection limits of 818 fM for the p.G12D and 1.8 pM for the wild-type targets. The genosensing assay was tested on genomic tumor DNA samples provided by IRCCS-Regina Elena National Cancer Institute and finally integrated into a smart portable multichannel potentiostat capable of performing up to four simultaneous acquisitions. The developed magneto-genosensing assay demonstrated portability, simplicity, and high sensitivity, showing good potential as theranostic tool for personalized medicine in oncology. |
| first_indexed | 2024-03-09T01:26:06Z |
| format | Article |
| id | doaj.art-23305f6eed1d44ebb5bfe7f1fb3282f1 |
| institution | Directory Open Access Journal |
| issn | 2590-1370 |
| language | English |
| last_indexed | 2024-03-09T01:26:06Z |
| publishDate | 2023-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Biosensors and Bioelectronics: X |
| spelling | doaj.art-23305f6eed1d44ebb5bfe7f1fb3282f12023-12-10T06:17:25ZengElsevierBiosensors and Bioelectronics: X2590-13702023-12-0115100404A highly sensitive electrochemical magneto-genosensing assay for the specific detection of a single nucleotide variation in the KRAS oncogene in human plasmaSimone Fortunati0Chiara Giliberti1Marco Giannetto2Alessandro Bertucci3Sabrina Capodaglio4Elena Ricciardi5Patrizio Giacomini6Valentina Bianchi7Andrea Boni8Ilaria De Munari9Roberto Corradini10Maria Careri11University of Parma, Department of Chemistry, Life Sciences and Environmental Sustainability, Parco Area Delle Scienze 17/A, 43124, Parma, ItalyUniversity of Parma, Department of Chemistry, Life Sciences and Environmental Sustainability, Parco Area Delle Scienze 17/A, 43124, Parma, ItalyUniversity of Parma, Department of Chemistry, Life Sciences and Environmental Sustainability, Parco Area Delle Scienze 17/A, 43124, Parma, Italy; Corresponding author.University of Parma, Department of Chemistry, Life Sciences and Environmental Sustainability, Parco Area Delle Scienze 17/A, 43124, Parma, ItalyUniversity of Parma, Department of Chemistry, Life Sciences and Environmental Sustainability, Parco Area Delle Scienze 17/A, 43124, Parma, ItalyUOC Oncological Translational Research, IRCCS-Regina Elena National Cancer Institute, 00144, Rome, ItalyClinical Trial Center, IRCSS-Regina Elena National Cancer Institute/Istituto Dermatologico San Gallicano, 00144, Rome, ItalyUniversity of Parma, Department of Engineering and Architecture, Parco Area Delle Scienze 181/A, 43124, Parma, ItalyUniversity of Parma, Department of Engineering and Architecture, Parco Area Delle Scienze 181/A, 43124, Parma, ItalyUniversity of Parma, Department of Engineering and Architecture, Parco Area Delle Scienze 181/A, 43124, Parma, ItalyUniversity of Parma, Department of Chemistry, Life Sciences and Environmental Sustainability, Parco Area Delle Scienze 17/A, 43124, Parma, ItalyUniversity of Parma, Department of Chemistry, Life Sciences and Environmental Sustainability, Parco Area Delle Scienze 17/A, 43124, Parma, ItalyLiquid biopsy is a non-invasive and efficient technique for the detection of tumor biomarkers in biological fluids, which currently represents a new frontier in theranostics and precision medicine. Among mutations of the KRAS oncogene, p.G12D single nucleotide variation in the KRAS gene plays a central role in the early diagnosis and therapeutic treatment of colorectal cancer. In this context, we developed a highly sensitive magneto-genosensing assay based on PNA capture probes immobilized on magnetic microbeads. To detect the mutation of the KRAS oncogene, two capture probe sequences recognizing wild-type and p.G12D tumor DNA were used in association with a DNA signaling probe allowing for electrochemical detection using an enzyme conjugate. The assay conditions were optimized using a 32 full-factorial design, obtaining an outstanding specificity, evidenced by a remarkably lower (>95%) signal of the singly-mismatched- compared to that of fully-complementary-DNA. Ultra-high sensitivity was achieved in 10-fold diluted human plasma reaching detection limits of 818 fM for the p.G12D and 1.8 pM for the wild-type targets. The genosensing assay was tested on genomic tumor DNA samples provided by IRCCS-Regina Elena National Cancer Institute and finally integrated into a smart portable multichannel potentiostat capable of performing up to four simultaneous acquisitions. The developed magneto-genosensing assay demonstrated portability, simplicity, and high sensitivity, showing good potential as theranostic tool for personalized medicine in oncology.http://www.sciencedirect.com/science/article/pii/S2590137023001012Point-of-CarePrecision medicineKRASColorectal cancerElectrochemical genoassayMagnetic microbeads |
| spellingShingle | Simone Fortunati Chiara Giliberti Marco Giannetto Alessandro Bertucci Sabrina Capodaglio Elena Ricciardi Patrizio Giacomini Valentina Bianchi Andrea Boni Ilaria De Munari Roberto Corradini Maria Careri A highly sensitive electrochemical magneto-genosensing assay for the specific detection of a single nucleotide variation in the KRAS oncogene in human plasma Biosensors and Bioelectronics: X Point-of-Care Precision medicine KRAS Colorectal cancer Electrochemical genoassay Magnetic microbeads |
| title | A highly sensitive electrochemical magneto-genosensing assay for the specific detection of a single nucleotide variation in the KRAS oncogene in human plasma |
| title_full | A highly sensitive electrochemical magneto-genosensing assay for the specific detection of a single nucleotide variation in the KRAS oncogene in human plasma |
| title_fullStr | A highly sensitive electrochemical magneto-genosensing assay for the specific detection of a single nucleotide variation in the KRAS oncogene in human plasma |
| title_full_unstemmed | A highly sensitive electrochemical magneto-genosensing assay for the specific detection of a single nucleotide variation in the KRAS oncogene in human plasma |
| title_short | A highly sensitive electrochemical magneto-genosensing assay for the specific detection of a single nucleotide variation in the KRAS oncogene in human plasma |
| title_sort | highly sensitive electrochemical magneto genosensing assay for the specific detection of a single nucleotide variation in the kras oncogene in human plasma |
| topic | Point-of-Care Precision medicine KRAS Colorectal cancer Electrochemical genoassay Magnetic microbeads |
| url | http://www.sciencedirect.com/science/article/pii/S2590137023001012 |
| work_keys_str_mv | AT simonefortunati ahighlysensitiveelectrochemicalmagnetogenosensingassayforthespecificdetectionofasinglenucleotidevariationinthekrasoncogeneinhumanplasma AT chiaragiliberti ahighlysensitiveelectrochemicalmagnetogenosensingassayforthespecificdetectionofasinglenucleotidevariationinthekrasoncogeneinhumanplasma AT marcogiannetto ahighlysensitiveelectrochemicalmagnetogenosensingassayforthespecificdetectionofasinglenucleotidevariationinthekrasoncogeneinhumanplasma AT alessandrobertucci ahighlysensitiveelectrochemicalmagnetogenosensingassayforthespecificdetectionofasinglenucleotidevariationinthekrasoncogeneinhumanplasma AT sabrinacapodaglio ahighlysensitiveelectrochemicalmagnetogenosensingassayforthespecificdetectionofasinglenucleotidevariationinthekrasoncogeneinhumanplasma AT elenaricciardi ahighlysensitiveelectrochemicalmagnetogenosensingassayforthespecificdetectionofasinglenucleotidevariationinthekrasoncogeneinhumanplasma AT patriziogiacomini ahighlysensitiveelectrochemicalmagnetogenosensingassayforthespecificdetectionofasinglenucleotidevariationinthekrasoncogeneinhumanplasma AT valentinabianchi ahighlysensitiveelectrochemicalmagnetogenosensingassayforthespecificdetectionofasinglenucleotidevariationinthekrasoncogeneinhumanplasma AT andreaboni ahighlysensitiveelectrochemicalmagnetogenosensingassayforthespecificdetectionofasinglenucleotidevariationinthekrasoncogeneinhumanplasma AT ilariademunari ahighlysensitiveelectrochemicalmagnetogenosensingassayforthespecificdetectionofasinglenucleotidevariationinthekrasoncogeneinhumanplasma AT robertocorradini ahighlysensitiveelectrochemicalmagnetogenosensingassayforthespecificdetectionofasinglenucleotidevariationinthekrasoncogeneinhumanplasma AT mariacareri ahighlysensitiveelectrochemicalmagnetogenosensingassayforthespecificdetectionofasinglenucleotidevariationinthekrasoncogeneinhumanplasma AT simonefortunati highlysensitiveelectrochemicalmagnetogenosensingassayforthespecificdetectionofasinglenucleotidevariationinthekrasoncogeneinhumanplasma AT chiaragiliberti highlysensitiveelectrochemicalmagnetogenosensingassayforthespecificdetectionofasinglenucleotidevariationinthekrasoncogeneinhumanplasma AT marcogiannetto highlysensitiveelectrochemicalmagnetogenosensingassayforthespecificdetectionofasinglenucleotidevariationinthekrasoncogeneinhumanplasma AT alessandrobertucci highlysensitiveelectrochemicalmagnetogenosensingassayforthespecificdetectionofasinglenucleotidevariationinthekrasoncogeneinhumanplasma AT sabrinacapodaglio highlysensitiveelectrochemicalmagnetogenosensingassayforthespecificdetectionofasinglenucleotidevariationinthekrasoncogeneinhumanplasma AT elenaricciardi highlysensitiveelectrochemicalmagnetogenosensingassayforthespecificdetectionofasinglenucleotidevariationinthekrasoncogeneinhumanplasma AT patriziogiacomini highlysensitiveelectrochemicalmagnetogenosensingassayforthespecificdetectionofasinglenucleotidevariationinthekrasoncogeneinhumanplasma AT valentinabianchi highlysensitiveelectrochemicalmagnetogenosensingassayforthespecificdetectionofasinglenucleotidevariationinthekrasoncogeneinhumanplasma AT andreaboni highlysensitiveelectrochemicalmagnetogenosensingassayforthespecificdetectionofasinglenucleotidevariationinthekrasoncogeneinhumanplasma AT ilariademunari highlysensitiveelectrochemicalmagnetogenosensingassayforthespecificdetectionofasinglenucleotidevariationinthekrasoncogeneinhumanplasma AT robertocorradini highlysensitiveelectrochemicalmagnetogenosensingassayforthespecificdetectionofasinglenucleotidevariationinthekrasoncogeneinhumanplasma AT mariacareri highlysensitiveelectrochemicalmagnetogenosensingassayforthespecificdetectionofasinglenucleotidevariationinthekrasoncogeneinhumanplasma |