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评估马乳头瘤、耳部斑块和肉瘤中是否存在马乳头瘤病毒DNA和乳头瘤病毒抗原。

Evaluation of equine papillomas, aural plaques, and sarcoids for the presence of Equine papillomavirus DNA and Papillomavirus antigen.

作者信息

Postey Rosemary C, Appleyard Greg D, Kidney Beverly A

机构信息

Department of Veterinary Pathology, Western College of Veterinary Medicine, University of Saskatchewan, 52 Campus Drive, Saskatoon, Saskatchewan S7N 5B4.

出版信息

Can J Vet Res. 2007 Jan;71(1):28-33.

Abstract

Immunohistochemical (IHC) testing and electron microscopy have implicated Papillomavirus (PV) as the etiologic agent for equine papillomas and aural plaques, but Equine papillomavirus (EPV) DNA has yet to be demonstrated in these lesions by polymerase chain reaction (PCR). The purpose of this study was to evaluate formalin-fixed, paraffin-embedded tissues from naturally occurring cases of equine papillomas, aural plaques, and sarcoids for the presence of EPV DNA by means of PCR and for the presence of PV antigen by means of IHC testing. We used EPV-specific primers that amplified a region of 384 base pairs (bp) spanning the E4 and L2 genes of the EPV genome and consensus PV primers that amplified a 102-bp region of the L1 gene. Group-specific PV structural antigens were detected with the use of a streptavidin-biotin-alkaline phosphatase IHC stain. With IHC testing, 23 of 38 papillomas, 4 of 9 aural plaques, and 0 of 10 sarcoids were positive for PV antigen; EPV DNA was found in 20 of the 38 papillomas and 1 of the 10 sarcoids but 0 of the 9 aural plaques. The consensus primers did not amplify novel PV DNA in any of the tissues. Nucleotide sequencing of viral DNA from 7 papillomas amplified with EPV-specific primers revealed DNA fragments that were 96% to 99% identical to known EPV sequences. Some samples had nucleotide substitutions in common, which suggests infection with related strains. Together, EPV DNA or PV antigen (or both) was demonstrated in 26 (68%) of the 38 equine papillomas. Although aural plaques contained PV antigen, they were negative for EPV DNA; therefore, we hypothesize that aural plaques contain a PV distinct from EPV.

摘要

免疫组织化学(IHC)检测和电子显微镜检查表明乳头瘤病毒(PV)是马乳头瘤和耳部斑块的病原体,但通过聚合酶链反应(PCR)尚未在这些病变中检测到马乳头瘤病毒(EPV)DNA。本研究的目的是通过PCR评估福尔马林固定、石蜡包埋的来自自然发生的马乳头瘤、耳部斑块和肉瘤病例的组织中EPV DNA的存在情况,并通过IHC检测评估PV抗原的存在情况。我们使用了EPV特异性引物,其扩增了跨越EPV基因组E4和L2基因的384个碱基对(bp)的区域,以及通用PV引物,其扩增了L1基因的102 bp区域。使用链霉亲和素-生物素-碱性磷酸酶IHC染色检测组特异性PV结构抗原。通过IHC检测,38个乳头瘤中的23个、9个耳部斑块中的4个和10个肉瘤中的0个PV抗原呈阳性;在38个乳头瘤中的20个和10个肉瘤中的1个中发现了EPV DNA,但9个耳部斑块中均未发现。通用引物在任何组织中均未扩增出新的PV DNA。对用EPV特异性引物扩增的7个乳头瘤的病毒DNA进行核苷酸测序,结果显示DNA片段与已知EPV序列的同一性为96%至99%。一些样本存在共同的核苷酸替换,这表明感染了相关毒株。38个马乳头瘤中有26个(68%)检测到了EPV DNA或PV抗原(或两者都有)。尽管耳部斑块含有PV抗原,但EPV DNA呈阴性;因此,我们推测耳部斑块含有一种不同于EPV的PV。

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