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@article{1798685,
author = {Izadi, Nasim and Sebuyoya, Ravery and Moráňová, Ludmila and Hrstka, Roman and Anton, Milan and Bartošík, Martin},
article_location = {Amsterdam},
article_number = {December 2021},
doi = {http://dx.doi.org/10.1016/j.aca.2021.339145},
keywords = {HPV; Cervical cancer; Crude lysate; Electrochemistry; LAMP amplification},
language = {eng},
issn = {0003-2670},
journal = {Analytica Chimica Acta},
title = {Electrochemical bioassay coupled to LAMP reaction for determination of high-risk HPV infection in crude lysates},
url = {https://www.sciencedirect.com/science/article/pii/S0003267021009715?via%3Dihub},
volume = {1187},
year = {2021}
}
TY - JOUR
ID - 1798685
AU - Izadi, Nasim - Sebuyoya, Ravery - Moráňová, Ludmila - Hrstka, Roman - Anton, Milan - Bartošík, Martin
PY - 2021
TI - Electrochemical bioassay coupled to LAMP reaction for determination of high-risk HPV infection in crude lysates
JF - Analytica Chimica Acta
VL - 1187
IS - December 2021
SP - 1-9
EP - 1-9
PB - Elsevier
SN - 00032670
KW - HPV
KW - Cervical cancer
KW - Crude lysate
KW - Electrochemistry
KW - LAMP amplification
UR - https://www.sciencedirect.com/science/article/pii/S0003267021009715?via%3Dihub
N2 - Electrochemical (EC) detection of DNA biomarkers represents an interesting tool in molecular oncology due to its sensitivity, simplicity, low cost or rapid times of measurement. However, majority of EC assays, same as most optical-based techniques, require preceding DNA extraction step to remove other cellular components, making these assays more laborious and time-consuming. One option to circumvent this is to use LAMP (loop-mediated amplification), an isothermal amplification technique that can amplify DNA directly in crude lysates in a short time at a constant temperature. Here, we coupled the LAMP reaction with EC readout to detect DNA from the two most common oncogenic human papillomavirus (HPV) types that cause cervical cancer in women, i.e. HPV 16 and HPV 18, directly in crude lysates without a need for DNA extraction step. We show that in crude lysates, the LAMP reaction was superior to PCR, with very good selectivity on a panel of cancer cell lines and with high sensitivity, enabling detection of HPV DNA from as few as 10 cells. As a proof of principle, we applied the assay to nineteen clinical samples both from uninfected women and from women suffering from cervical precancerous lesions caused by HPV 16 or HPV 18 genotypes. Clinical samples were simply boiled for 5 min in homogenization buffer without DNA extraction step, and amplified with LAMP. We obtained excellent concordance of our assay with PCR, reaching 100% sensitivity for both genotypes, 81.82% specificity for HPV 16 and 94.12% spec-ificity for HPV 18. Proposed assay could be a straightforward, simple, rapid and sensitive alternative for early diagnostics of precancerous cervical lesions.
ER -
IZADI, Nasim, Ravery SEBUYOYA, Ludmila MORÁŇOVÁ, Roman HRSTKA, Milan ANTON and Martin BARTOŠÍK. Electrochemical bioassay coupled to LAMP reaction for determination of high-risk HPV infection in crude lysates. \textit{Analytica Chimica Acta}. Amsterdam: Elsevier, 2021, vol.~1187, December 2021, p.~1-9. ISSN~0003-2670. Available from: https://dx.doi.org/10.1016/j.aca.2021.339145.
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