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

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00209805%3A_____%2F21%3A00078877" target="_blank" >RIV/00209805:_____/21:00078877 - isvavai.cz</a>

Alternative codes found

RIV/65269705:_____/21:00075827 RIV/00216224:14310/21:00122608

Result on the web

<a href="https://pubmed.ncbi.nlm.nih.gov/34753575/" target="_blank" >https://pubmed.ncbi.nlm.nih.gov/34753575/</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1016/j.aca.2021.339145" target="_blank" >10.1016/j.aca.2021.339145</a>

Result language

angličtina

Original language name

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

Original language description

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% specificity for HPV 18. Proposed assay could be a straightforward, simple, rapid and sensitive alternative for early diagnostics of precancerous cervical lesions.

Czech name

—

Czech description

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Type

J_imp - Article in a specialist periodical, which is included in the Web of Science database

CEP classification

—

OECD FORD branch

10405 - Electrochemistry (dry cells, batteries, fuel cells, corrosion metals, electrolysis)

Project

Result was created during the realization of more than one project. More information in the Projects tab.

Continuities

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Publication year

2021

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

Analytica chimica acta

ISSN

0003-2670

e-ISSN

—

Volume of the periodical

1187

Issue of the periodical within the volume

December

Country of publishing house

NL - THE KINGDOM OF THE NETHERLANDS

Number of pages

9

Pages from-to

339145.

UT code for WoS article

000718364300003

EID of the result in the Scopus database

2-s2.0-85116866898