Genomagnetic LAMP-based electrochemical test for determination of high-risk HPV16 and HPV18 in clinical samples

Multiplexed Immunosensing Platform Coupled to Hybridization Chain Reaction for Electrochemical Determination of MicroRNAs in Clinical Samples

“…Some of the experimental conditions: the dilution of AbS9.6 (1 : 250 in PBS), temperature of reactions (37 °C), ratio of hydroquinone/hydrogen peroxide (10 mM/50 mM), and applied potential (−0.3 V/60 s) for the chronoamperometric detection, were optimized in our previous works . With the aim of developing a methodology that allows the sensitive determination of the target miRNA with a short and simple working protocol, the number of incubation steps was carefully evaluated.…”

“…We focused on cervical cancer on purpose. Recently, we have developed an EC‐based assay for detection of DNA from HPV, the virus causing virtually all cases of this type of cancer, and applied it successfully to real patient samples . Although current HPV testing (to which our published assay also belongs) establishes itself as an important alternative to traditional cytological examination of the cervical specimen (called the Pap test), reaching better sensitivities and thus decreasing number of false negatives, it does not provide information on the virus activity but only on its presence.…”

“…Moreover, the efficient incorporation of the resulting magnetic bioconjugates on the surface of planar disposable electrodes in a stable and reproducible way by simple magnetic attraction also implies that detectable products, usually generated through an enzyme reaction, are formed very close to the electrode surface, thus allowing the electrochemical response to be obtained rapidly . In addition, it is important to remark that although the coupling of MBs with single or dual disposable electrodes has been widely described, their use with a screen‐printed carbon electrode array of eight electrodes has been reported only recently by our group . Three miRNAs, miR‐21, let‐7a and miR‐31, associated with development and progression of cervical cancer in women , were selected as target miRNAs.…”

Loop-mediated isothermal amplification (LAMP) – review and classification of methods for sequence-specific detection

“…The insertion of biotin into amplicons by biotin-labelled primers is unproblematic regarding the efficiency of amplication due to Aedes aegypti (Wolbachia infection) 85,86 Ammonia-oxidizing enzyme in environmental bacteria 64 Avian reovirus 70 Bacteriophage MS2 73 Bemisia tabaci 60 BRAF allele (V600E) 83 Brucella 72 Caenorhabditis elegans 68 Chikungunya virus 73 Chlamydia trachomatis 61 Dengue virus 87,112,156 Diarrheal disease 82 Escherichia coli 68 Fomitiporia torreyae 59 Fulviformes umbrinellus 59 Fusarium oxysporum f. sp. lycopersici (point mutations in xylem 3 (SIX3) gene) 66 Genetically modied maize [77][78][79][80][81] Haemophilus ducreyi 101 Haemophilus inuenzae 105 hBRCA1 68 HeLa 68 Hepatitis B and C virus 62,87,107 Herpes simplex virus 1 (HSV1) US4 83 Human immunodeciency virus 87,101 HLA-B*15:02 allele 163 Human papillomavirus 153,171 Human T-lymphotropic virus 1 101 Inuenza virus 63,114 Lambda DNA 68 Leptospira 148 Leishmania 155 Listeria ivanovii 103 Listeria monocytogenes 103 Methicillin-resistant Staphylococcus aureus (MRSA) 71,166 Middle east respiratory sy...…”

“…The turnaround time including LAMP is about 2.5 hours. This method was presented by Bartosik et al 171,172 for the detection of HPV types 16 and 18 on electrochemical chips using a multi potentiostat/galvanostat. The analytical sensitivity was described with a limit of detection of 0.1 ng DNA per reaction and a limit of quantication of 0.6 ng per reaction.…”