Evaluation of 16S rRNA Hypervariable Regions for Bioweapon Species Detection by Massively Parallel Sequencing
Joint Authors
Dias, Victor H. G.
Gomes, Priscila da S. F. C.
Azevedo-Martins, Allan C.
Cabral, Bianca C. A.
Woerner, August E.
Budowle, Bruce
Moura-Neto, Rodrigo S.
Silva, Rosane
Source
International Journal of Microbiology
Issue
Vol. 2020, Issue 2020 (31 Dec. 2020), pp.1-11, 11 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2020-09-26
Country of Publication
Egypt
No. of Pages
11
Main Subjects
Abstract EN
Molecular detection and classification of the bacterial groups in a sample are relevant in several areas, including medical research and forensics.
Sanger sequencing of the 16S rRNA gene is considered the gold standard for microbial phylogenetic analysis.
However, the development of massively parallel sequencing (MPS) offers enhanced sensitivity and specificity for microbiological analyses.
In addition, 16S rRNA target amplification followed by MPS facilitates the combined use of multiple markers/regions, better discrimination of sample background, and higher sample throughput.
We designed a novel set of 16S rRNA gene primers for detection of bacterial species associated with clinical, bioweapon, and biohazards microorganisms via alignment of 364 sequences representing 19 bacterial species and strains relevant to medical and forensics applications.
In silico results indicated that the hypervariable regions (V1V2), (V4V5), and (V6V7V8) support the resolution of a selected group of bacteria.
Interspecies and intraspecies comparisons showed 74.23%–85.51% and 94.48%–99.98% sequencing variation among species and strains, respectively.
Sequence reads from a simulated scenario of bacterial species mapped to each of the three hypervariable regions of the respective species with different affinities.
The minimum limit of detection was achieved using two different MPS platforms.
This protocol can be used to detect or monitor as low as 2,000 genome equivalents of bacterial species associated with clinical, bioweapon, and biohazard microorganisms and potentially can distinguish natural outbreaks of pathogenic microorganisms from those occurring by intentional release.
American Psychological Association (APA)
Dias, Victor H. G.& Gomes, Priscila da S. F. C.& Azevedo-Martins, Allan C.& Cabral, Bianca C. A.& Woerner, August E.& Budowle, Bruce…[et al.]. 2020. Evaluation of 16S rRNA Hypervariable Regions for Bioweapon Species Detection by Massively Parallel Sequencing. International Journal of Microbiology،Vol. 2020, no. 2020, pp.1-11.
https://search.emarefa.net/detail/BIM-1172461
Modern Language Association (MLA)
Dias, Victor H. G.…[et al.]. Evaluation of 16S rRNA Hypervariable Regions for Bioweapon Species Detection by Massively Parallel Sequencing. International Journal of Microbiology No. 2020 (2020), pp.1-11.
https://search.emarefa.net/detail/BIM-1172461
American Medical Association (AMA)
Dias, Victor H. G.& Gomes, Priscila da S. F. C.& Azevedo-Martins, Allan C.& Cabral, Bianca C. A.& Woerner, August E.& Budowle, Bruce…[et al.]. Evaluation of 16S rRNA Hypervariable Regions for Bioweapon Species Detection by Massively Parallel Sequencing. International Journal of Microbiology. 2020. Vol. 2020, no. 2020, pp.1-11.
https://search.emarefa.net/detail/BIM-1172461
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1172461