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Microbiology Time – March 2024

Microbiology Time

This month, we bring you a new oral microbiota sampling method, a study on saliva and Long COVID, and yet another paper highlighting the importance of self-collection in the fight against cervical cancer.

  • This first study investigated the association between COVID-19 and Long COVID in children. The Italian researchers conducted a prospective study involving 167 pediatric patients hospitalized with confirmed acute SARS-CoV-2 infection, and collecting saliva samples with LolliSponge. A telephone survey was conducted at 3-6-12 months after discharge. Of the patients, 95.5% presented with symptoms upon admission. Regarding patients for whom it was feasible to determine the SARS-CoV-2 variant, the study identified the Delta variant in 21.6% and the Omicron variant in 78.4%. About 16.5% of patients reported symptoms indicative of Long COVID, such as weight loss, inappetence, chronic cough, fatigue, and others. Only 4 patients with Long COVID had a specific SARS-CoV-2 variant identified. The study emphasizes the significant concern of Long COVID in the pediatric population, highlighting the need for continuous monitoring and tailored symptom investigation based on age.
  • For the second study, we remain in Italy. Marianna Martinelli and colleagues assessed the accuracy of the BD Onclarity HPV assay on self-collected vaginal FLOQSwabs compared to cervical samples for detecting cervical intraepithelial neoplasia (CIN). Testing was conducted on two automated platforms, BD Viper LT and BD COR, and included 300 women. Self-sampling was performed with two vaginal swabs to assess, with the second one, the influence of freezing on the results. The results showed similar sensitivity for ≥CIN2 between vaginal and cervical samples, regardless of the platform used. Specificity was initially slightly lower for self-collected samples compared to cervical samples, but cut-off optimization for HPV positivity improved relative specificity without compromising sensitivity. No significant differences were observed between self-samples tested with the two platforms and between first- and second-collected (frozen) swabs. The study concludes that Onclarity assay on self-collected FLOQSwabs samples showed similar sensitivity and specificity (with cut-off optimization) to detect ≥CIN2 compared to cervical samples.
  • The third study addressed the challenge of standardizing oral microbiome analysis due to the variability in microbiota compositions across different oral habitats and the lack of standardized collection methods. The study proposes a standardized protocol for dental examination and material collection for next-generation sequencing (NGS) from various oral sites. The researchers involved 60 patients grouped into COVID-19 convalescents with and without antibiotics and healthy individuals. The collected biological samples were then used to conduct NGS (16S rRNA). Alpha and beta diversity analyses reveal notable differences in microbial communities, particularly in the supragingival plaque. The study emphasizes the importance of precise procedures during dental exams and material collection to avoid technical errors and contamination in NGS results. Dental indices like DMFT, D number, PI, and BOP are highlighted as valuable prognostic parameters for assessing oral health. The developed procedures are suggested for use in future studies by future studies.

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Microbiology Time – February 2024

Microbiology Time

This month we dig into salivary micro-RNA as a liquid biopsy marker in oral cancer, different methods of offering HPV self-sampling kits, and the benefits of probiotics in improving vaginal microbiota composition in post-menopausal women.

  • The first study investigates the use of salivary cell-free miRNA as a liquid biopsy marker in cancer. The researchers focused on establishing reproducible methods for saliva manipulation to control pre-analytical variables affecting miRNA stability. Saliva samples from healthy subjects and oral cancer patients were collected using two methods (spit and LolliSponge), and storage conditions were varied. Quantifying the salivary expression level of target miRNAs by qPCR, the study found comparable levels between the two collection methods. MiRNAs remained stable for up to 48 hours at 4 ◦C but showed significant alterations after 96 hours. Mid-term storage at -20 ◦C decreased miRNA stability compared to -80 ◦C. The study concludes that cell-free miRNA in saliva, though slightly affected by collection methods and storage conditions, remains stable for a duration compatible with clinical routine needs.
  • In the second study, the US researchers performed a randomized clinical trial to assess the effectiveness of different approaches for offering HPV self-sampling kits to individuals based on their cervical cancer screening history (screening-adherent and currently due, overdue, or unknown). The individuals were stratified to receive usual care, education, direct mail with a self-sampling kit, and an opt-in option. The results also showed that direct-mail self-sampling significantly increased cervical cancer screening by over 14% in individuals who were due or overdue. At the same time, the opt-in approach had minimal impact. The study suggests prioritizing direct-mail outreach for those due or overdue for screening to enhance adherence, while for individuals with unknown screening history, alternative outreach approaches and efforts to document screening history are recommended.
  • The third study is a prospective observational clinical trial conducted in Italy, aiming at investigating the benefits of probiotics in improving vaginal well-being and microbiota composition in post-menopausal women. The study enrolled 50 healthy post-menopausal women who took a supplement containing specific probiotic strains for 28 days. Results showed a decrease in menopausal symptoms, with significant improvements in the Vaginal Health Index (VHI) score by 50%, a reduction in inflammatory cytokine levels, and restoration of vaginal microbiota, including an increase in lactobacilli abundance. The study suggests that the tested probiotic strains, previously effective in childbearing-age women, are also beneficial for post-menopausal women, demonstrating improvements in both vaginal well-being and microbiota composition.

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