Electrochemical DNA biosensor coupled to LAMP reaction for early diagnostics of cervical precancerous lesions

Sebuyoya, Ravery; Moráňová, Ludmila; Izadi, Nasim; Moráň, Lukáš; Hrstka, Roman; Anton, Milan; Bartošík, Martin

doi:10.1016/j.biosx.2022.100224

Cited by 5 publications

(5 citation statements)

References 39 publications

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Section: Isothermal Amplification Techniques As Rapid Alternatives To...mentioning

confidence: 99%

“…Our team has published several research articles reporting fast and simple LAMP‐based assays combined with electrochemical detection (EC‐LAMP), targeting HPV16/18 genotypes in both cervical cell lines and cervical smears from women with precancerous lesions (Figure 1). 79–82 After development and optimization of the assay, 80 the EC‐LAMP was applied into a cohort of 61 clinical samples to evaluate its performance and compared it with PCR‐based methods and with INNO‐LiPA genotyping assay 81 . Good specificity and negative and positive predictive values were reached, all over 90%.…”

Section: Isothermal Amplification Techniques As Rapid Alternatives To...mentioning

confidence: 99%

“…68 Our team has published several research articles reporting fast and simple LAMP-based assays combined with electrochemical detection (EC-LAMP), targeting HPV16/18 genotypes in both cervical cell lines and cervical smears from women with precancerous lesions (Figure 1). [79][80][81][82] After development and optimization of the assay, 80…”

Section: Electrochemical (Ec) End-point Detection Is Another Suitablementioning

confidence: 99%

“…This elimination not only simplified overall assay, but also decreased a risk of contamination by shortening overall sample exposition to the environment. The last generation of the assay involved a transfer of the protocol from magnetic beads directly to the gold electrode chips for easier washing and shorter hands-on time 82. LAMP as a rapid and undemanding technique could be thus a good choice for fast determination of HPV infection.Another IAT, the recombinase polymerase amplification (RPA), often functions with forward and reverse PCR primers, but instead of a Taq polymerase, it uses a cocktail of three enzymes-recombinase (enabling primers to pair with homologous sequences in target DNA strand), single-stranded DNA-binding protein (stabilizing singlestranded DNA structures and preventing displacement of primers), and strand-displacing polymerase (performing actual synthesis of complementary DNA).…”

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confidence: 99%

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Advanced technologies towards improved HPV diagnostics

Bartosik,

Moranova,

Izadi

et al. 2024

Journal of Medical Virology

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Persistent infection with high‐risk types of human papillomaviruses (HPV) is a major cause of cervical cancer, and an important factor in other malignancies, for example, head and neck cancer. Despite recent progress in screening and vaccination, the incidence and mortality are still relatively high, especially in low‐income countries. The mortality and financial burden associated with the treatment could be decreased if a simple, rapid, and inexpensive technology for HPV testing becomes available, targeting individuals for further monitoring with increased risk of developing cancer. Commercial HPV tests available in the market are often relatively expensive, time‐consuming, and require sophisticated instrumentation, which limits their more widespread utilization. To address these challenges, novel technologies are being implemented also for HPV diagnostics that include for example, isothermal amplification techniques, lateral flow assays, CRISPR‐Cas‐based systems, as well as microfluidics, paperfluidics and lab‐on‐a‐chip devices, ideal for point‐of‐care testing in decentralized settings. In this review, we first evaluate current commercial HPV tests, followed by a description of advanced technologies, explanation of their principles, critical evaluation of their strengths and weaknesses, and suggestions for their possible implementation into medical diagnostics.

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Section: Isothermal Amplification Techniques As Rapid Alternatives To...mentioning

confidence: 99%

Section: Isothermal Amplification Techniques As Rapid Alternatives To...mentioning

confidence: 99%

Section: Electrochemical (Ec) End-point Detection Is Another Suitablementioning

confidence: 99%

mentioning

confidence: 99%

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Advanced technologies towards improved HPV diagnostics

Bartosik,

Moranova,

Izadi

et al. 2024

Journal of Medical Virology

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“…Upraveno dle cit. 14 nímu paralelnímu sekvenování nové generace bylo identifikováno velké množství kódujících i nekódujících RNA, jejichž hladina se mění během vzniku nebo progrese nádorového onemocnění. Mezi známé příklady kódujících biomarkerů, tj.…”

Section: Rna Nádorové Biomarkeryunclassified

Electrochemical Analysis of Clinical Samples – A Path to Translational Research

Moráňová,

Bartošík,

Strmisková

et al. 2023

Chem. Listy

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Precise diagnostics of cancer or other diseases is crucial when selecting proper treatment. Personalized medicine puts high demands on the accuracy of nucleic acid biomarkers analysis, where subtle differences at the nucleotide level are often involved. Isothermal amplification techniques offer new possibilities of DNA and RNA amplification without using PCR, and their combination with electrochemistry provide a promising fast and cost-effective alternative diagnostic tool. Although electrochemical biosensors are still insufficiently applied to clinical material, thus hindering their development, recent advancements show great promise in translational research. Banks of biological material (biobanks) are specialized workplaces focused on the long-term preservation and processing of clinical material and offer a wide range of expert services, primarily for research purposes, in particular the provision of biological samples and associated pseudonymized data. Their involvement in the field of electrochemical biosensors can facilitate application of electrochemical methods into clinical laboratories and expand the portfolio of currently used diagnostic methods.

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Electrochemical Detection of BRAF V600E Mutation in a Liquid Biopsy Format: LAMPing the Way to an Improved Diagnostics and Treatment

Moranova,

Strmiskova,

Ondraskova

et al. 2024

Adv Materials Technologies

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The BRAF V600E substitution, predominantly found in melanoma and metastatic colorectal cancer (mCRC), is associated with aggressive traits and poor prognosis. Molecular testing of the BRAF mutational status is crucial for guiding therapeutic decisions, driving the necessity for new rapid diagnostic technologies. Here, a novel LAMP‐based assay is presented for fast and selective amplification of mutated DNA, coupled with simple electrochemical detection on electrode biochips. The assay demonstrated high selectivity across cell lines and in 51 cancer patients, detecting mutations in both tumor tissue DNA and circulating tumor DNA (ctDNA) in serum. Clear discrimination between the mutated and wild‐type sequences is achieved, in perfect agreement with either NGS or ddPCR. The bioassay offers a simple and efficient alternative to traditional PCR methods for point‐of‐care diagnostics, particularly in settings with limited equipment and resources.

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Electrochemical DNA biosensor coupled to LAMP reaction for early diagnostics of cervical precancerous lesions

Cited by 5 publications

References 39 publications

Advanced technologies towards improved HPV diagnostics

Advanced technologies towards improved HPV diagnostics

Electrochemical Analysis of Clinical Samples – A Path to Translational Research

Electrochemical Detection of BRAF V600E Mutation in a Liquid Biopsy Format: LAMPing the Way to an Improved Diagnostics and Treatment

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