bioRxiv Science⌕ Search

Biology subjects

Robinson, C. K.

Publications and source records attributed to Robinson, C. K..

2 recordsLinked to original sources

Comparative metagenome-assembled genome analysis of Lachnovaginosum genomospecies, formerly known as BVAB1

Bacterial Vaginosis Associated Bacteria 1 (BVAB1) is an uncultured bacterial species found in the human vagina that belongs to the family Lachnospiraceae within the order Clostridiales, and as its name suggests, is often associated with bacterial vaginosis (BV). In reproductive health, BV is of concern due to its associated risk for HIV, Chlamydia trachomatis, and Neisseria gonorrhoeae acquisition as well as preterm birth. BVAB1 has been shown to be associated with BV persistence after metronidazole treatment and increased vaginal inflammation, and to confer a higher risk of puerperal infections. To date, no genome of BVAB1 is available, which has made it difficult to understand its disease-associated features. We present here a comparative analysis of seven metagenome-assembled genomes (MAGs) of BVAB1, derived from the cervicovaginal lavages of seven separate women. One metagenome was sequenced with long-read technology on a PacBio Sequel II instrument while the others were sequenced using the Illumina HiSeq platform. The MAGs were 1.5-1.7 Mb long, encoding an average of 1,499 genes and had >98% average nucleotide identity to each other. CheckM-estimated genome completion was 98.2%. We propose to rename BVAB1 to Lachnovaginosum genomospecies based on a phylogenetic analysis, and provide genomic evidence that this species may metabolize D-lactate, produce trimethylamine (the chemical responsible for BV-associated odor), and be motile. These MAGs will be a valuable resource and will contribute to our understanding of the heterogenous etiologies of bacterial vaginosis.

microbiology↗

Ultra-high throughput multiplexing and sequencing of >500 bp amplicon regions on the Illumina HiSeq 2500 platform

Amplification, sequencing and analysis of the 16S rRNA gene affords characterization of microbial community composition. As this tool has become more popular and amplicon-sequencing applications have grown in the total number of samples, growth in sample multiplexing is becoming necessary while maintaining high sequence quality and sequencing depth. Here, modifications to the Illumina HiSeq 2500 platform are described which produce greater multiplexing capabilities and 300 bp paired-end reads of higher quality than produced by the current Illumina MiSeq platform. To improve the feasibility and flexibility of this method, a 2-Step PCR amplification protocol is also described that allows for targeting of different amplicon regions, thus improving amplification success from low bacterial bioburden samples.\n\nImportanceAmplicon sequencing has become a popular and widespread tool for surveying microbial communities. Lower overall costs associated with high throughput sequencing have made it a widely-adopted approach, especially for projects which necessitate sample multiplexing to eliminate batch effect and reduced time to acquire data. The method for amplicon sequencing on the Illumina HiSeq 2500 platform described here provides improved multiplexing capabilities while simultaneously producing greater quality sequence data and lower per sample cost relative to the Illumina MiSeq platform, without sacrificing amplicon length. To make this method more flexible to various amplicon targeted regions as well as improve amplification from low biomass samples, we also present and validate a 2-Step PCR library preparation method.

microbiology↗