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Parkinson, N. J.

Publications and source records attributed to Parkinson, N. J..

2 recordsLinked to original sources

Long-read shotgun metagenomics identifies distinct urogenital microbial community states associated with functional potential and urinary tract infection during pregnancy

Urinary tract infections (UTIs) are among the most common bacterial infections during pregnancy and are associated with adverse maternal and neonatal outcomes. Although previous studies used enhanced culture, 16S rRNA gene sequencing, and/or short-read shotgun metagenomic sequencing to characterize the urogenital microbiome of adult females, few have assessed the urogenital microbiome during pregnancy and thus this microbiome remains poorly understood. Here, we applied Oxford Nanopore long-read shotgun metagenomic sequencing to characterize urogenital microbial composition, functional potential, metagenome-assembled genomes (MAGs), and the urogenital virome of pregnant females. Taxonomic profiling identified three distinct microbial community states predominated by Lactobacillus crispatus, Gardnerella species, and L. iners. The Gardnerella-predominated community was associated with a higher frequency of UTI during pregnancy. Functional profiles strongly reflected these community states, demonstrating that taxonomic differences were accompanied by distinct functional potential. Genome-resolved analysis recovered 415 MAGs, including 29 high-quality genomes, revealing substantial genomic diversity among urogenital microbes. Viral sequences, predominantly bacteriophages, also were detected in most samples, highlighting an additional component of the pregnancy-associated urogenital microbiome. These findings demonstrate that pregnant females possess distinct urogenital microbial community states and show that they differ not only in taxonomic composition but also in their functional potential.

microbiology↗

A comparison of two culture and three sequencing methods for detection of the urobiome

Standard urine culture (SUC), the clinical reference method, is optimized for fast-growing aerobes and misses much of the urinary microbiota. Enhanced culture and sequencing methods have rarely been compared on the same specimens. We compared five methods (SUC, expanded quantitative urine culture [EQUC], short-read V4 16S rRNA gene sequencing, long-read near full-length 16S rRNA gene sequencing, and PCR-free long-read shotgun metagenomic sequencing) on parallel aliquots of urine from pregnant participants screened for asymptomatic bacteriuria. Of 98 cultured samples, 98 (100.0%) were EQUC-positive while 85 (86.7%) SUC-positive. EQUC detected 102 species, but SUC only detected 41. Short-read 16S rRNA gene sequencing, long-read 16S rRNA gene sequencing, and shotgun sequencing were positive for 188/188, 186/188 and 139/139 samples, detecting 195, 69 and 528 species, respectively. In the 79 samples positive by EQUC and all three sequencing methods, sequencing profiles resembled each other more than culture profiles, and SUC captured only a subset of taxa detected by every other method. Shotgun sequencing gave the broadest taxonomic coverage and was the only sequencing method to identify Escherichia coli to the species level. With EQUC, it outperformed both 16S sequencing methods in detecting Gardnerella vaginalis and distinguishing Lactobacillus species, and it resolved six Gardnerella species, three detected by no other method. Long-read 16S rRNA gene sequencing gave the most confident amplicon-based species assignments, whereas culture better recovered Corynebacterium and Staphylococcus species. No single method was best across all endpoints, but long-read shotgun metagenomics provided the broadest species-level view. Method choice should match the research or clinical question; outcome-based studies are needed to establish clinical utility.

bioinformatics↗