Silver-enhanced in situ hybridization for detection of polyomavirus DNA in patients with BK virus nephropathy.

Florian R Fritzsche, Silvio Pianca, Ariana Gaspert, Zsuzsanna Varga, Lin Wang, Michael P Farrell, Xiao-Bo Chen, Hans H Hirsch, Erik Springer, Thomas Fehr, Jonathan Myles, Raymond Tubbs, Holger Moch
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引用次数: 4

Abstract

BK virus nephropathy is not an infrequent complication of renal transplantation associated with high rates of graft loss. Although antibodies against SV40 antigen detect different viruses of the polyomavirus family, immunohistochemistry is widely used to confirm the diagnosis of BK virus nephropathy in renal biopsies. Here we aimed to validate the novel silver-enhanced in situ hybridization (SISH) technique for the automated detection of BK virus in renal transplant biopsies. Two different patient cohorts were included. Twenty-nine consecutive patients suspicious for BK virus infection were investigated by SISH and chromogenic in situ hybridization. An additional 26 renal biopsies positive by SV40 immunohistochemistry from 19 patients were analyzed by SISH. Polyomavirus DNA serum levels, as determined by nested PCR analysis, were available for all of these patients. The presence of BK virus DNA in renal tubular cells was identified in 5 of the suspicious cases by both, SISH and chromogenic in situ hybridization . One additional patient was only positive in the SISH. In the second cohort, SISH was positive in all SV40 positive biopsies, but SISH signals were less extensive than SV40 immunohistochemistry. Our results show that the BK virus SISH is an ancillary tool for the detection of polyomavirus DNA in renal biopsies using bright-field microscopy. However, its diagnostic value in comparison with standard immunohistochemistry seems to be limited.

银增强原位杂交检测BK病毒肾病患者多瘤病毒DNA。
BK病毒肾病并不是肾移植的罕见并发症,它与移植肾丢失率高有关。虽然针对SV40抗原的抗体检测多瘤病毒家族的不同病毒,但免疫组织化学在肾活检中被广泛用于确认BK病毒肾病的诊断。在这里,我们旨在验证新的银增强原位杂交(SISH)技术在肾移植活检中自动检测BK病毒。纳入了两个不同的患者队列。对29例疑似BK病毒感染的患者进行了SISH和显色原位杂交检测。另外19例患者的26例肾活检呈SV40免疫组化阳性,用SISH分析。通过巢式PCR分析确定的多瘤病毒DNA血清水平可用于所有这些患者。5例可疑病例肾小管细胞均存在BK病毒DNA,经SISH和显色原位杂交鉴定。另有1例患者仅在SISH中呈阳性。在第二组队列中,SISH在所有SV40阳性活检中均呈阳性,但SISH信号的广泛程度不及SV40免疫组化。我们的研究结果表明,BK病毒SISH是一种辅助工具,用于检测多瘤病毒DNA在肾活检使用明场显微镜。然而,与标准免疫组织化学相比,其诊断价值似乎有限。
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来源期刊
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审稿时长
>12 weeks
期刊介绍: Diagnostic Molecular Pathology focuses on providing clinical and academic pathologists with coverage of the latest molecular technologies, timely reviews of established techniques, and papers on the applications of these methods to all aspects of surgical pathology and laboratory medicine. It publishes original, peer-reviewed contributions on molecular probes for diagnosis, such as tumor suppressor genes, oncogenes, the polymerase chain reaction (PCR), and in situ hybridization. Articles demonstrate how these highly sensitive techniques can be applied for more accurate diagnosis.
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