Evaluation of 16S rRNA primer sets for characterisation of microbiota in paediatric patients with autism spectrum disorder

Abstract Abstract intestinal microbiota is becoming a significant marker that reflects differences between health and disease status also in terms of gut-brain axis communication. Studies show that children with autism spectrum disorder (ASD) often have a mix of gut microbes that is distinct from th...

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Autores principales:	L. Palkova, A. Tomova, G. Repiska, K. Babinska, B. Bokor, I. Mikula, G. Minarik, D. Ostatnikova, K. Soltys
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/013366bbfc27401ba8c6f2b47c70265a
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spelling	oai:doaj.org-article:013366bbfc27401ba8c6f2b47c70265a2021-12-02T17:04:34ZEvaluation of 16S rRNA primer sets for characterisation of microbiota in paediatric patients with autism spectrum disorder10.1038/s41598-021-86378-w2045-2322https://doaj.org/article/013366bbfc27401ba8c6f2b47c70265a2021-03-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-86378-whttps://doaj.org/toc/2045-2322Abstract Abstract intestinal microbiota is becoming a significant marker that reflects differences between health and disease status also in terms of gut-brain axis communication. Studies show that children with autism spectrum disorder (ASD) often have a mix of gut microbes that is distinct from the neurotypical children. Various assays are being used for microbiota investigation and were considered to be universal. However, newer studies showed that protocol for preparing DNA sequencing libraries is a key factor influencing results of microbiota investigation. The choice of DNA amplification primers seems to be the crucial for the outcome of analysis. In our study, we have tested 3 primer sets to investigate differences in outcome of sequencing analysis of microbiota in children with ASD. We found out that primers detected different portion of bacteria in samples especially at phylum level; significantly higher abundance of Bacteroides and lower Firmicutes were detected using 515f/806r compared to 27f/1492r and 27f*/1495f primers. So, the question is whether a gold standard of Firmicutes/Bacteroidetes ratio is a valuable and reliable universal marker, since two primer sets towards 16S rRNA can provide opposite information. Moreover, significantly higher relative abundance of Proteobacteria was detected using 27f/1492r. The beta diversity of sample groups differed remarkably and so the number of observed bacterial genera.L. PalkovaA. TomovaG. RepiskaK. BabinskaB. BokorI. MikulaG. MinarikD. OstatnikovaK. SoltysNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-13 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q L. Palkova A. Tomova G. Repiska K. Babinska B. Bokor I. Mikula G. Minarik D. Ostatnikova K. Soltys Evaluation of 16S rRNA primer sets for characterisation of microbiota in paediatric patients with autism spectrum disorder
description	Abstract Abstract intestinal microbiota is becoming a significant marker that reflects differences between health and disease status also in terms of gut-brain axis communication. Studies show that children with autism spectrum disorder (ASD) often have a mix of gut microbes that is distinct from the neurotypical children. Various assays are being used for microbiota investigation and were considered to be universal. However, newer studies showed that protocol for preparing DNA sequencing libraries is a key factor influencing results of microbiota investigation. The choice of DNA amplification primers seems to be the crucial for the outcome of analysis. In our study, we have tested 3 primer sets to investigate differences in outcome of sequencing analysis of microbiota in children with ASD. We found out that primers detected different portion of bacteria in samples especially at phylum level; significantly higher abundance of Bacteroides and lower Firmicutes were detected using 515f/806r compared to 27f/1492r and 27f*/1495f primers. So, the question is whether a gold standard of Firmicutes/Bacteroidetes ratio is a valuable and reliable universal marker, since two primer sets towards 16S rRNA can provide opposite information. Moreover, significantly higher relative abundance of Proteobacteria was detected using 27f/1492r. The beta diversity of sample groups differed remarkably and so the number of observed bacterial genera.
format	article
author	L. Palkova A. Tomova G. Repiska K. Babinska B. Bokor I. Mikula G. Minarik D. Ostatnikova K. Soltys
author_facet	L. Palkova A. Tomova G. Repiska K. Babinska B. Bokor I. Mikula G. Minarik D. Ostatnikova K. Soltys
author_sort	L. Palkova
title	Evaluation of 16S rRNA primer sets for characterisation of microbiota in paediatric patients with autism spectrum disorder
title_short	Evaluation of 16S rRNA primer sets for characterisation of microbiota in paediatric patients with autism spectrum disorder
title_full	Evaluation of 16S rRNA primer sets for characterisation of microbiota in paediatric patients with autism spectrum disorder
title_fullStr	Evaluation of 16S rRNA primer sets for characterisation of microbiota in paediatric patients with autism spectrum disorder
title_full_unstemmed	Evaluation of 16S rRNA primer sets for characterisation of microbiota in paediatric patients with autism spectrum disorder
title_sort	evaluation of 16s rrna primer sets for characterisation of microbiota in paediatric patients with autism spectrum disorder
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/013366bbfc27401ba8c6f2b47c70265a
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Evaluation of 16S rRNA primer sets for characterisation of microbiota in paediatric patients with autism spectrum disorder

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