gastro-jРоссийский журнал гастроэнтерологии, гепатологии, колопроктологииRussian Journal of Gastroenterology, Hepatology, Coloproctology1382-43762658-6673«Gastro» LLCgastro-j-1730Research ArticleОРИГИНАЛЬНЫЕ ИССЛЕДОВАНИЯORIGINAL ARTICLESОпределение микробной флоры пигментных желчных камней на основе анализа гена 16S рибосомальной РНКInvestigation of microbial flora of pigment gallstones on the basis of 16S ribosomal RNA gene analysisЛаринА. К.LarinA. K.ЩербаковП. Л.ScherbakovP. L.ХаритоноваЛ. А.KharitonovaL. A.ЦарьковаО. А.TsarkovaO. A.МомыналиевК. Т.MomynaliyevK. T.2009051020091954954Copyright © Ларин А.К., Щербаков П.Л., Харитонова Л.А., Царькова О.А., Момыналиев К.Т., 20092009Ларин А.К., Щербаков П.Л., Харитонова Л.А., Царькова О.А., Момыналиев К.Т.Larin A.K., Scherbakov P.L., Kharitonova L.A., Tsarkova O.A., Momynaliyev K.T.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.gastro-j.ru/jour/article/view/1730

Цель исследования. Оценка микробной флоры пигментных желчных камней с помощью анализа гена 16S рибосомальной РНК.Материал и методы. При холецистэктомии у детей, инфицированных H. pylori, взято 7 образцов пигментных желчных камней. Для идентификации микроорганизмов использовали метод клональных библиотек. С помощью ПЦР амплифицировали участок гена 16S рибосомальной РНК, осуществляли лигирование, трансформацию E. coli и получали библиотеки клонов. Определяли нуклеотидную последовательность фрагмента гена 16S рРНК в не менее чем 100 клонах. Проводили компьютерный анализ полученной информации и устанавливали принадлежность прочитанных нуклеотидных последовательностей тому или иному микроорганизму с помощью базы данных NCBI.Результаты. В 4 образцах черных пигментных камней бактериальной ДНК не обнаружено. Для 3 образцов коричневых пигментных камней созданы библиотеки фрагментов гена 16S рРНК, определены их последовательности и идентифицированы микроорганизмы, которым они принадлежат.Заключение. У детей, инфицированных H. pylori, этот микроб не является причиной формирования желчных камней.

Aim of investigation

Aim of investigation. Investigation of microbial flora of pigment gallstones by analysis of ribosomal 16S RNA gene.

Material and methods

Material and methods. Seven samples of pigment gallstones were obtained at cholecystectomy from children infected by H. pylori. DNA amplification method (PCR) was used for identification of microorganisms. Ligation, E. coli transformation and creation of clonal libraries were carried out. Nucleotide sequence was determined. Computer analysis of the obtained information was done; belonging of the read nucleotide sequences to certain microorganism by NCBI database was accomplished.

Results

Results. In 4 samples of black pigment stones no bacterial DNA was revealed. In 3 samples of brown pigment stones libraries of 16S rRNA gene fragments were created, their sequences were determined and microorganisms to which they belong were identified.

Conclusion

Conclusion. At children infected by H. pylori, this microbe is not the cause of gallstones development.

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The authors declare that there are no conflicts of interest present.