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

Kód výsledku v 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>

Nalezeny alternativní kódy

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

Výsledek na webu

<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>

Jazyk výsledku

angličtina

Název v původním jazyce

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

Popis výsledku v původním jazyce

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.

Název v anglickém jazyce

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

Popis výsledku anglicky

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.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

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

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

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

Rok uplatnění

2021

Kód důvěrnosti údajů

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

Název periodika

Analytica chimica acta

ISSN

0003-2670

e-ISSN

—

Svazek periodika

1187

Číslo periodika v rámci svazku

December

Stát vydavatele periodika

NL - Nizozemsko

Počet stran výsledku

9

Strana od-do

339145.

Kód UT WoS článku

000718364300003

EID výsledku v databázi Scopus

2-s2.0-85116866898