Detailed Information on Publication Record
2021
Electrochemical bioassay coupled to LAMP reaction for determination of high-risk HPV infection in crude lysates
IZADI, Nasim, Ravery SEBUYOYA, Ludmila MORÁŇOVÁ, Roman HRSTKA, Milan ANTON et. al.Basic information
Original name
Electrochemical bioassay coupled to LAMP reaction for determination of high-risk HPV infection in crude lysates
Authors
IZADI, Nasim (364 Islamic Republic of Iran, guarantor), Ravery SEBUYOYA (834 Tanzania, United Republic of, belonging to the institution), Ludmila MORÁŇOVÁ (203 Czech Republic, belonging to the institution), Roman HRSTKA (203 Czech Republic), Milan ANTON (203 Czech Republic, belonging to the institution) and Martin BARTOŠÍK (703 Slovakia)
Edition
Analytica Chimica Acta, Amsterdam, Elsevier, 2021, 0003-2670
Other information
Language
English
Type of outcome
Článek v odborném periodiku
Field of Study
10406 Analytical chemistry
Country of publisher
Netherlands
Confidentiality degree
není předmětem státního či obchodního tajemství
References:
Impact factor
Impact factor: 6.911
RIV identification code
RIV/00216224:14310/21:00122608
Organization unit
Faculty of Science
UT WoS
000718364300003
Keywords in English
HPV; Cervical cancer; Crude lysate; Electrochemistry; LAMP amplification
Tags
International impact, Reviewed
Změněno: 16/5/2022 11:57, Mgr. Marie Šípková, DiS.
Abstract
V originále
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.
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