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Quantitative determination of beta-lactamase mRNA in the RNA-transcripts of antibiotic-resistant bacteria using colorimetric biochipsстатья
Информация о цитировании статьи получена из
Scopus
Статья опубликована в журнале из списка Web of Science и/или Scopus
Дата последнего поиска статьи во внешних источниках: 8 июня 2022 г.
- Авторы: Rubtsova М.Yu, Filippova A.A., Fursova N.K., Grigorenko V.G., Presnova G.V., Ulyashova M.M., Egorov А.М.
- Журнал: Journal of Analytical Chemistry
- Том: 77
- Номер: 5
- Год издания: 2022
- Издательство: Maik Nauka/Interperiodica Publishing
- Местоположение издательства: Russian Federation
- Первая страница: 519
- Последняя страница: 530
- DOI: 10.1134/S1061934822050124
- Аннотация: A method for the quantitative determination of mRNA of TEM-type serine beta-lactamases inbacteria resistant to beta-lactam antibiotics has been developed. The method includes several stages: the isolation of a total RNA fraction from a bacterial culture and the production of the beta-lactamase target DNA in sequential reverse transcription and polymerase chain reaction with the incorporation of a biotin label, andthe hybridization of the target DNA on colorimetric biochips. Sample preparation conditions were optimized to increase the yield of the analyzed target DNA. A calibration curve for determining the amount of mRNA was constructed using a standard sample of beta-lactamase TEM-1 mRNA obtained by transcription in vitro. The advantage of using a standard sample corresponding to the full-length blaTEM-1 gene is that it passes through all the stages of analysis in parallel with the test samples with the same efficiency. The detection limit for TEM-1 beta-lactamase mRNA was 0.40 ± 0.05 amol/mL; the range of determined oncentrations was from 1.0 amol/mL to 2000 fmol/mL, and the relative standard deviation did not exceed 12%. The duration of analysis after the production of a bacterial culture was about 7 h. The developed method can be used to study the conditions for the expression of beta-lactamase genes in bacteria resistant to antimicrobial drugs.
- Добавил в систему: Рубцова Майя Юрьевна
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