High-sensitivity BK virus detection system using viewRNA in situ hybridization

IF 2.1 4区医学 Q3 INFECTIOUS DISEASES

Diagnostic microbiology and infectious disease Pub Date : 2025-03-05 DOI:10.1016/j.diagmicrobio.2025.116790

Chunlan Hu , Xiaonan Zhang , Tongyu Zhu , Yumin Hou , Yejing Shi , Jiajia Sun , Nannan Wu

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引用次数: 0

Abstract

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.

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采用viewRNA原位杂交技术的高灵敏度BK病毒检测系统

目的建立可靠的BKV相关病理分子诊断系统。方法设计了一套针对BKV大T抗原区两个保守序列的原位杂交（ISH）探针，对JC病毒和SV40同源物具有高度特异性。通过将分支DNA信号扩增技术与这些探针相结合，我们开发了一种新的ViewRNA-ISH检测系统。结果优化后的BKV-LT ViewRNA-ISH系统在细胞模型和临床标本中均比常规免疫组织化学方法具有更高的诊断特异性和敏感性。29个临床样本和3个皮下植入肿瘤样本的验证显示，IHC结果与IHC结果的一致性为93.75%(30/32)，其中2个先前IHC阴性的肾活检通过ViewRNA-ISH检测显示阳性信号。结论BKV- lt ViewRNA-ISH系统可实现BKV核酸双模式（荧光/化学）检测，对BKV相关病变具有较好的诊断效果，在同种异体肾移植监测中具有潜在的临床应用价值。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Diagnostic microbiology and infectious disease 医学-传染病学

CiteScore

5.30

自引率

3.40%

发文量

149

审稿时长

56 days

期刊介绍： Diagnostic Microbiology and Infectious Disease keeps you informed of the latest developments in clinical microbiology and the diagnosis and treatment of infectious diseases. Packed with rigorously peer-reviewed articles and studies in bacteriology, immunology, immunoserology, infectious diseases, mycology, parasitology, and virology, the journal examines new procedures, unusual cases, controversial issues, and important new literature. Diagnostic Microbiology and Infectious Disease distinguished independent editorial board, consisting of experts from many medical specialties, ensures you extensive and authoritative coverage.