TTV Load Mirrors Local Immunity and Tracks HPV Positivity in the Anogenital Tract

ABSTRACT Torquetenovirus load has been proposed as an immunomodulated biomarker of host immune status, yet its behavior in situ within HPV‐infected mucosa remains poorly defined. We conducted a retrospective cross‐sectional study of 220 patients undergoing HPV screening (181 cervical swabs, 39 anal brushings). HPV was genotyped with Allplex™ HPV28, and TTV load was quantified by in‐house RT‐qPCR and expressed as log 10 copies per 20 ng total DNA. Analyses included nonparametric group comparisons (Mann‐Whitney) and site‐stratified logistic regression to estimate the TTV‐HPV association. Genotype co‐occurrence was summarized by heatmaps and network analysis and formally tested with Fisher's exact test with Benjamini‐Hochberg FDR correction. TTV load was higher in HPV‐positive subjects ( p < 0.01), with a significant difference in BR and a trend in CS. In a binary logistic model, each 1‐log 10 increase in TTV was associated with a 60% increase in the odds of HPV positivity (OR = 1.60, 95% CI 1.07–2.39; p = 0.022), with consistent results across CS and BR. TTV load increased with greater genotypic complexity in co‐infections (genotype richness) and was also higher in infections sustained exclusively by HR genotypes than by LR genotypes ( p < 0.01). The co‐occurrence map and network analysis highlighted recurrent genotype combinations (e.g., 16/53, 53/68, 42/53) and central nodes (16, 31, 51, 56, 68, 53). In situ quantification of TTV is associated with HPV positivity and with genotype complexity/risk class, offering local predictive power. The co‐occurrence evidence remains exploratory but supports TTV as an indirect indicator of mucosal immunocompetence.