High-sensitivity BK virus detection system using viewRNA in situ hybridization
Chunlan Hu1, Xiaonan Zhang2, Tongyu Zhu3
1Shanghai Public Health Clinical Center, Fudan University, Shanghai 201508, China; Department of Kidney Transplantation, Zhongshan Hospital, Fudan University, Shanghai 200032, China.
Objective:
To establish a reliable molecular diagnostic system for precise identification of BK virus (BKV)-associated pathologies.
Methods:
We designed a set of in-situ hybridization (ISH) probes targeting two conserved sequences within the large T antigen region of BKV, exhibiting highly specificity from JC virus and SV40 homologs. By integrating the branched DNA signal amplification technology with these probes, we developed a novel ViewRNA-ISH detection system.
Results:
The optimized BKV-LT ViewRNA-ISH system demonstrated enhanced diagnostic specificity and sensitivity compared to conventional immunohistochemical methods in both cellular models and clinical specimens. Validation using 29 clinical samples and 3 subcutaneous planted tumor samples revealed 93.75 % (30/32) concordance with IHC findings, with 2 previously IHC-negative renal biopsies showing positive signals through ViewRNA-ISH detection.
Conclusions:
Our BKV-LT ViewRNA-ISH system enables dual-mode (fluorescent/ chemical) detection of BKV nucleic acids, providing superior diagnostic performance for BKV-associated pathologies with potential clinical utility in renal allograft monitoring.
