Multiplex detection in tonsillar tissue of all known human polyomaviruses

Area of Science:

• Virology
• Immunology
• Molecular Biology

Background:

• The Polyomaviridae family includes 13 known human polyomaviruses (HPyVs), with infections common since childhood.
• While widespread, the persistence sites and transmission routes of HPyVs are largely unknown.
• Previous research suggested lymphoid tissue, specifically tonsils, as a potential persistence site for novel HPyVs.

Purpose of the Study:

• To develop and validate a Luminex-based multiplex assay for simultaneous detection of all 13 HPyVs.
• To investigate the prevalence and distribution of HPyVs in human tonsillar tissues from children and adults.

Main Methods:

• A Luminex-based multiplex assay was designed and validated using primer pairs and probes targeting the HPyV VP1 gene.
• The assay demonstrated high analytical sensitivity (10^0-10^2 copies/µL) and specificity, with no cross-reactivity observed.
• 78 tonsillar tissue samples were tested for the presence of DNA from all 13 HPyVs.

Main Results:

• The multiplex assay successfully detected all 13 HPyVs simultaneously with high accuracy.
• HPyV DNA was detected in 14 out of 78 (17.9%) tonsillar samples.
• The most frequently detected HPyVs were HPyV6 (7.7%), TSPyV (3.8%), and WUPyV (3.8%); one sample showed mixed infection.

Conclusions:

• The Luminex-based HPyV multiplex assay is suitable for clinical diagnostics and epidemiological studies.
• Findings provide further evidence for the role of the lymphoid system in HPyV infection and persistence.
• Tonsillar shedding during HPyV reactivation may contribute to the transmission of newly discovered HPyVs.