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Record W4415666730 · doi:10.1038/s41598-025-21406-7

Real-time PCR assay design and validation for Prevotella bivia, Peptostreptococcus anaerobius, and Dialister micraerophilus, bacteria associated with increased HIV susceptibility

2025· article· en· W4415666730 on OpenAlexafffund
Sydney G. Nelson, Megan Spyers-Duran, Maliha Aziz, Danielle Marcone, Deneen Nault, Gizachew Demessie, Rupert Kaul, Ronald M. Galiwango, Rebecca Yee, Cindy M. Liu

Bibliographic record

VenueScientific Reports · 2025
Typearticle
Languageen
FieldBiochemistry, Genetics and Molecular Biology
TopicBacterial Identification and Susceptibility Testing
Canadian institutionsUniversity of Toronto
FundersCanadian Institutes of Health ResearchFogarty International CenterNational Institutes of HealthGeorge Washington University
KeywordsPeptostreptococcusPrevotellaBacteriaGardnerella vaginalisAnaerobic bacteriaImmune systemIn vitro

Abstract

fetched live from OpenAlex

Many genital anaerobes, including Prevotella bivia , Peptostreptococcus anaerobius , and Dialister micraerophilus , are underexplored despite being Bacteria Associated with HIV Seroconversion, Inflammation, and immune Cells (BASICs). This is partly due to the lack of molecular tools for their detection and quantification. To address this gap, we designed and validated three real-time qPCR assays for rapid and cost-effective analysis. qPCR assays were designed based on the species-specific core genomes of the three target genital anaerobic species. Assay sensitivity, specificity, and quantification characteristics were assessed using a synthetic oligonucleotide and DNA extracted from closely related species ( n = 27–42) and human urogenital swabs ( n = 111–114). The resultant assays demonstrated 100% sensitivity and specificity for bacterial isolates and high sensitivity (94.6%-97.7%) and specificity (92.8%-95.7%) for human urogenital swabs. The linear dynamic range was 5.0 × 10 1 to 1.0 × 10 7 copies/µL for P. bivia and D. micraerophilus assays, and 2.5 × 10 2 to 1.0 × 10 7 copies/µL for the P. anaerobius assay. Assay efficiency ranged from 99.0% to 112.0%. These assays provide a highly sensitive and specific method for the analysis of bacterial isolates, urogenital samples, and in vitro or ex vivo experiments, which can enhance our understanding of the epidemiology, clinical impact, and biology of key genital anaerobes associated with HIV risk.

Fetched live from OpenAlex and de-inverted. Abstracts are not stored in this database: the inverted indexes are 8.6 GB of the frame’s 9.3 GB of text, and the host has 13 GB free.

How this classification was reachedexpand

Full frame machine prediction

Teacher imitation

Not calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.

metaresearch head score (Codex)0.009
metaresearch head score (Gemma)0.010
Version: metacan-v3-hybrid-931329e0061cValidation status: machine_predicted_unvalidated
Candidate categoriesnone
Consensus categoriesnone
DomainCandidate signal: none · Consensus signal: none
Study designCandidate signal: Bench or experimental · Consensus signal: Bench or experimental
GenreCandidate signal: Methods · Consensus signal: Methods
Teacher disagreement score0.009
Threshold uncertainty score0.049

Distilled classifier scores by category (both heads)

CategoryCodexGemma
Metaresearch0.0090.010
Meta-epidemiology (narrow)0.0020.002
Meta-epidemiology (broad)0.0020.002
Bibliometrics0.0020.001
Science and technology studies0.0010.002
Scholarly communication0.0020.001
Open science0.0020.001
Research integrity0.0020.002
Insufficient payload (model declined to judge)0.0010.001

Machine scores (provisional)

The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.

Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.

Opus teacher head0.013
GPT teacher head0.246
Teacher spread0.233 · how far apart the two teachers sit on this one work
Validation statusscore_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from it

Classification

machine, unvalidated

Machine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.

The models applied no category: nothing in the taxonomy fit this work.
Study designBench or experimental
Domainnot available
GenreMethods

How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".

Quick stats

Citations1
Published2025
Admission routes2
Has abstractyes

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