Untangling Species-Level Composition of Complex Bacterial Communities through a Novel Metagenomic Approach
ABSTRACT 16S small-subunit (SSU) rRNA gene-based bacterial profiling is the gold standard for cost-effective taxonomic reconstruction of complex bacterial populations down to the genus level. However, it has been proven ineffective in clinical and research settings requiring higher taxonomic resolut...
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American Society for Microbiology
2020
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oai:doaj.org-article:d4e2d1baae8347529ba54dc3171c60772021-12-02T18:15:46ZUntangling Species-Level Composition of Complex Bacterial Communities through a Novel Metagenomic Approach10.1128/mSystems.00404-202379-5077https://doaj.org/article/d4e2d1baae8347529ba54dc3171c60772020-08-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSystems.00404-20https://doaj.org/toc/2379-5077ABSTRACT 16S small-subunit (SSU) rRNA gene-based bacterial profiling is the gold standard for cost-effective taxonomic reconstruction of complex bacterial populations down to the genus level. However, it has been proven ineffective in clinical and research settings requiring higher taxonomic resolution. We therefore developed a bacterial profiling method based on the internal transcribed spacer (ITS) region employing optimized primers and a comprehensive ITS database for accurate cataloguing of bacterial communities at (sub)species resolution. Performance of the microbial ITS profiling pipeline was tested through analysis of host-associated, food, and environmental matrices, while its efficacy in clinical settings was assessed through analysis of mucosal biopsy specimens of colorectal cancer, leading to the identification of putative novel biomarkers. The data collected indicate that the proposed pipeline represents a major step forward in cost-effective identification and screening of microbial biomarkers at (sub)species level, with relevant impact in research, industrial, and clinical settings. IMPORTANCE We developed a novel method for accurate cataloguing of bacterial communities at (sub)species level involving amplification of the internal transcribed spacer (ITS) region through optimized primers, followed by next-generation sequencing and taxonomic classification of amplicons by means of a comprehensive database of bacterial ITS sequences. Host-associated, food, and environmental matrices were employed to test the performance of the microbial ITS profiling pipeline. Moreover, mucosal biopsy samples from colorectal cancer patients were analyzed to demonstrate the scientific relevance of this profiling approach in a clinical setting through identification of putative novel biomarkers. The results indicate that the ITS-based profiling pipeline proposed here represents a key metagenomic tool with major relevance for research, industrial, and clinical settings.Christian MilaniGiulia AlessandriMarta MangifestaLeonardo MancabelliGabriele Andrea LugliFederico FontanaGiulia LonghiRosaria AnzaloneAlice ViappianiSabrina DurantiFrancesca TurroniRenato CostiAlfredo AnnicchiaricoAndrea MoriniLeopoldo SarliMaria Cristina OssiprandiDouwe van SinderenMarco VenturaAmerican Society for MicrobiologyarticlemetagenomicsITSprofilingMicrobiologyQR1-502ENmSystems, Vol 5, Iss 4 (2020) |
| institution |
DOAJ |
| collection |
DOAJ |
| language |
EN |
| topic |
metagenomics ITS profiling Microbiology QR1-502 |
| spellingShingle |
metagenomics ITS profiling Microbiology QR1-502 Christian Milani Giulia Alessandri Marta Mangifesta Leonardo Mancabelli Gabriele Andrea Lugli Federico Fontana Giulia Longhi Rosaria Anzalone Alice Viappiani Sabrina Duranti Francesca Turroni Renato Costi Alfredo Annicchiarico Andrea Morini Leopoldo Sarli Maria Cristina Ossiprandi Douwe van Sinderen Marco Ventura Untangling Species-Level Composition of Complex Bacterial Communities through a Novel Metagenomic Approach |
| description |
ABSTRACT 16S small-subunit (SSU) rRNA gene-based bacterial profiling is the gold standard for cost-effective taxonomic reconstruction of complex bacterial populations down to the genus level. However, it has been proven ineffective in clinical and research settings requiring higher taxonomic resolution. We therefore developed a bacterial profiling method based on the internal transcribed spacer (ITS) region employing optimized primers and a comprehensive ITS database for accurate cataloguing of bacterial communities at (sub)species resolution. Performance of the microbial ITS profiling pipeline was tested through analysis of host-associated, food, and environmental matrices, while its efficacy in clinical settings was assessed through analysis of mucosal biopsy specimens of colorectal cancer, leading to the identification of putative novel biomarkers. The data collected indicate that the proposed pipeline represents a major step forward in cost-effective identification and screening of microbial biomarkers at (sub)species level, with relevant impact in research, industrial, and clinical settings. IMPORTANCE We developed a novel method for accurate cataloguing of bacterial communities at (sub)species level involving amplification of the internal transcribed spacer (ITS) region through optimized primers, followed by next-generation sequencing and taxonomic classification of amplicons by means of a comprehensive database of bacterial ITS sequences. Host-associated, food, and environmental matrices were employed to test the performance of the microbial ITS profiling pipeline. Moreover, mucosal biopsy samples from colorectal cancer patients were analyzed to demonstrate the scientific relevance of this profiling approach in a clinical setting through identification of putative novel biomarkers. The results indicate that the ITS-based profiling pipeline proposed here represents a key metagenomic tool with major relevance for research, industrial, and clinical settings. |
| format |
article |
| author |
Christian Milani Giulia Alessandri Marta Mangifesta Leonardo Mancabelli Gabriele Andrea Lugli Federico Fontana Giulia Longhi Rosaria Anzalone Alice Viappiani Sabrina Duranti Francesca Turroni Renato Costi Alfredo Annicchiarico Andrea Morini Leopoldo Sarli Maria Cristina Ossiprandi Douwe van Sinderen Marco Ventura |
| author_facet |
Christian Milani Giulia Alessandri Marta Mangifesta Leonardo Mancabelli Gabriele Andrea Lugli Federico Fontana Giulia Longhi Rosaria Anzalone Alice Viappiani Sabrina Duranti Francesca Turroni Renato Costi Alfredo Annicchiarico Andrea Morini Leopoldo Sarli Maria Cristina Ossiprandi Douwe van Sinderen Marco Ventura |
| author_sort |
Christian Milani |
| title |
Untangling Species-Level Composition of Complex Bacterial Communities through a Novel Metagenomic Approach |
| title_short |
Untangling Species-Level Composition of Complex Bacterial Communities through a Novel Metagenomic Approach |
| title_full |
Untangling Species-Level Composition of Complex Bacterial Communities through a Novel Metagenomic Approach |
| title_fullStr |
Untangling Species-Level Composition of Complex Bacterial Communities through a Novel Metagenomic Approach |
| title_full_unstemmed |
Untangling Species-Level Composition of Complex Bacterial Communities through a Novel Metagenomic Approach |
| title_sort |
untangling species-level composition of complex bacterial communities through a novel metagenomic approach |
| publisher |
American Society for Microbiology |
| publishDate |
2020 |
| url |
https://doaj.org/article/d4e2d1baae8347529ba54dc3171c6077 |
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