Electrochemical bioassay coupled to LAMP reaction for determination of high-risk HPV infection in crude lysates

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

doi:10.1016/j.aca.2021.339145

Cited by 22 publications

(19 citation statements)

References 34 publications

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“…Several researchers have reported high sensitivity and specificity HPV detection using LAMP reaction. But these LAMP assays took more than 60 min to get results, which cannot fulfill the gap for on-site detection. , We selected the E 1 and E 7 genes as markers for HPV 16 and HPV 18, respectively, and optimized real-time LAMP in bulk solutions to maximize the reaction speed, meanwhile minimizing nonspecific background amplification. The bulk solution LAMP optimization process consisted of four steps: (1) screening primer sets for high-speed and low-background amplification, (2) testing the FIP and BIP primers with a range of concentrations, (3) optimizing the LAMP solution with a range of magnesium ion concentrations, and (4) selecting the optimal amplification temperature.…”

Section: Resultsmentioning

confidence: 99%

Rapid In Situ Hydrogel LAMP for On-Site Large-Scale Parallel Single-Cell HPV Detection

et al. 2022

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Rapid human papillomavirus (HPV) screening is urgently needed for preventing and early diagnosis of cervical cancer in rural areas. To date, no HPV nucleic acid test (NAT) can be implemented within a single patient visit starting from clinical samples. Here, we develop a hydrogel loop-mediated isothermal amplification (LAMP) method in a fashion of large-scale parallel (about 1000 cells) in situ HPV DNA detection in clinical cervical exfoliated cells at the single-cell level. It can be used with a hotplate and smartphone to obtain HPV NAT results in less than 30 min, which is especially suitable for the on-site scenario. We apply this rapid HPV NAT on 40 clinical cervical exfoliated cell samples and compare the results to a clinical gold standard quantitative polymerase chain reaction (qPCR) method [area under curve (AUC), 1.00]. Meanwhile, our assay can provide HPV infection information for large-scale parallel single clinical cervical exfoliated cells, which cannot be received from traditional NAT methods. Our findings suggest the potential of in situ hydrogel LAMP as a powerful tool for clinical HPV screening and fundamental research.

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Section: Resultsmentioning

confidence: 99%

Rapid In Situ Hydrogel LAMP for On-Site Large-Scale Parallel Single-Cell HPV Detection

et al. 2022

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“…It requires 4-6 primers which makes the reaction highly sequence-specific and generates products of variable lengths, ranging from hundreds to few thousand bp. LAMP is widely used for pathogen analysis not only in a fluorescent or colorimetric format [117][118][119][120], but also in electrochemical assays to detect viruses, such as human papillomavirus (HPV) [121][122][123][124] and hepatitis virus B [125], and bacteria (E. coli, Salmonella, etc.) [126,127].…”

Section: Iat-based Ec Biosensorsmentioning

confidence: 99%

Electrochemical biosensors for analysis of DNA point mutations in cancer research

et al. 2022

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Cancer is a genetic disease induced by mutations in DNA, in particular point mutations in important driver genes that lead to protein malfunctioning and ultimately to tumorigenesis. Screening for the most common DNA point mutations, especially in such genes as TP53, BRCA1 and BRCA2, EGFR, KRAS, or BRAF, is crucial to determine predisposition risk for cancer or to predict response to therapy. In this review, we briefly depict how these genes are involved in cancer, followed by a description of the most common techniques routinely applied for their analysis, including high-throughput next-generation sequencing technology and less expensive low-throughput options, such as real-time PCR, restriction fragment length polymorphism, or high resolution melting analysis. We then introduce benefits of electrochemical biosensors as interesting alternatives to the standard methods in terms of cost, speed, and simplicity. We describe most common strategies involved in electrochemical biosensing of point mutations, relying mostly on PCR or isothermal amplification techniques, and critically discuss major challenges and obstacles that, until now, prevented their more widespread application in clinical settings.

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“…Izandi et al [80] demonstrated the relationship between cervical cancer and human Papilloma virus (HPV) by using the LAMP isothermal amplification technique coupled with an electrochemical detection platform. Specifically, they focused on the detection of HPV16 and HPV18, using a protocol that involves (1) lysis of tumor cells, (2) amplification by the LAMP reaction of virus DNA, (3) hybridization of the products on modified magnetic beads with specific capture probes, and (4) monitoring of the amplification reaction by the electrochemical detection.…”

Section: Isothermal Amplification Techniques Coupled To Electrochemic...mentioning

confidence: 99%

Chip-Based and Wearable Tools for Isothermal Amplification and Electrochemical Analysis of Nucleic Acids

et al. 2022

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The determination of nucleic acids has become an analytical diagnostic method with many applications in fields such as biomedical sciences, environmental monitoring, forensic identification, and food safety. Among the different methods for nucleic acid analysis, those based on the polymerase chain reaction (PCR) are nowadays considered the gold standards. Isothermal amplification methods are an interesting alternative, especially in the design of chip-based architectures. Biosensing platforms hold great promise for the simple and rapid detection of nucleic acids since they can be embedded in lab-on-a-chip tools to perform nucleic acid extraction, amplification, and detection steps. Electrochemical transduction schemes are particularly interesting in the design of small and portable devices due to miniaturization, low-energy consumption, and multianalyte detection capability. The aim of this review is to summarize the different applications of isothermal amplification methods combined with electrochemical biosensing techniques in the development of lab-on-a-chip tools and wearable sensors. Different isothermal amplification methods are revised, and examples of different applications are discussed. Finally, a discussion on patented devices is also included.

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Electrochemical bioassay coupled to LAMP reaction for determination of high-risk HPV infection in crude lysates

Cited by 22 publications

References 34 publications

Rapid In Situ Hydrogel LAMP for On-Site Large-Scale Parallel Single-Cell HPV Detection

Rapid In Situ Hydrogel LAMP for On-Site Large-Scale Parallel Single-Cell HPV Detection

Electrochemical biosensors for analysis of DNA point mutations in cancer research

Chip-Based and Wearable Tools for Isothermal Amplification and Electrochemical Analysis of Nucleic Acids

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