Multiplex detection in tonsillar tissue of all known human polyomaviruses

• Published in: BMC infectious diseases Vol. 17; no. 1; p. 409
• Main Authors: Sadeghi, Mohammadreza, Wang, Yilin, Ramqvist, Torbjörn, Aaltonen, Leena-Maija, Pyöriä, Lari, Toppinen, Mari, Söderlund-Venermo, Maria, Hedman, Klaus
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 08.06.2017; BioMed Central; BMC
• Subjects: Activation; Adolescent; Adult; Adults; Aged; Assaying; Biopsy; Capsid Proteins - genetics; Child; Child, Preschool; Children; Deoxyribonucleic acid; Detection limits; Diagnosis; Diagnostic systems; Disease; DNA; DNA probes; Epidemiology; Female; Gene sequencing; Genetic aspects; Genomes; Genotype; Health aspects; HPyV; Humans; Hypertrophy; Hypertrophy - diagnosis; Hypertrophy - epidemiology; Hypertrophy - pathology; Hypertrophy - virology; Infectious diseases; Innovations; Luminex; Lymphoid tissue; Male; Medicin och hälsovetenskap; Middle Aged; Mixed infection; Molecular diagnostic techniques; Multiplex Polymerase Chain Reaction - methods; Multiplexing; Nucleotide sequence; Palatine Tonsil - pathology; Palatine Tonsil - virology; Patients; PCR; Polyoma virus; Polyomavirus - genetics; Polyomavirus - isolation & purification; Polyomavirus Infections - diagnosis; Polyomavirus Infections - epidemiology; Polyomavirus Infections - pathology; Polyomavirus Infections - virology; Prevalence; Probes; Reproducibility of Results; Sensitivity analysis; Sensitivity and Specificity; Serology; Shedding; Target recognition; Throat surgery; Tissues; Tonsil; Tonsillitis; Tonsillitis - diagnosis; Tonsillitis - epidemiology; Tonsillitis - pathology; Tonsillitis - virology; Transplants & implants; Viruses; VP1 protein; Young Adult; HPyV; Tonsil; Luminex; PCR
• ISSN: 1471-2334; 1471-2334
• DOI: 10.1186/s12879-017-2479-5

In the past few years, eleven new human viruses have joined the two previously known members JCPyV and BKPyV of the Polyomaviridae family, by virtue of molecular methods. Serology data suggest that infections with human polyomaviruses (HPyVs) occur since childhood and the viruses are widespread in the general population. However, the viral persistence sites and transmission routes are by and large unknown. Our previous studies demonstrated that the four new HPyVs - KIPyV, WUPyV, MCPyV and TSPyV - were present in the tonsils, and suggested lymphoid tissue as a persistent site of these emerging human viruses. We developed a Luminex-based multiplex assay for simultaneous detection of all 13 HPyVs known, and explored their occurrence in tonsillar tissues of children and adults mostly with tonsillitis or tonsillar hypertrophy. We set up and validated a new Luminex-based multiplex assay by using primer pairs and probes targeting the respective HPyV viral protein 1 (VP1) genes. With this assay we tested 78 tonsillar tissues for DNAs of 13 HPyVs. The multiplex assay allowed for simultaneous detection of 13 HPyVs with high analytical sensitivity and specificity, with detection limits of 10 -10 copies per microliter, and identified correctly all 13 target sequences with no cross reactions. HPyV DNA altogether was found in 14 (17.9%) of 78 tonsils. The most prevalent HPyVs were HPyV6 (7.7%), TSPyV (3.8%) and WUPyV (3.8%). Mixed infection of two HPyVs occurred in one sample. The Luminex-based HPyV multiplex assay appears highly suitable for clinical diagnostic purposes and large-scale epidemiological studies. Additional evidence was acquired that the lymphoid system plays a role in HPyV infection and persistence. Thereby, shedding from this site during reactivation might take part in transmission of the newly found HPyVs.