Mazzei Maurizio, Forzan Mario, Carlucci Vito, Anfossi Antonio G, Alberti Alberto, Albanese Francesco, Binanti Diana, Millanta Francesca, Baroncini Lisa, Pirone Andrea, Abramo Francesca
1 Department of Veterinary Sciences, University of Pisa, Pisa, Italy.
2 Department of Veterinary Medicine, University of Sassari, Sassari, Italy.
J Feline Med Surg. 2018 Aug;20(8):772-779. doi: 10.1177/1098612X17732255. Epub 2017 Sep 26.
Objectives The aim of the study was to investigate, by quantitative PCR (qPCR), the presence of papillomavirus in feline viral plaques (VPs), Bowenoid in situ carcinoma (BISC), squamous cell carcinoma (SCC) and actinic keratosis (AK). Methods Twenty-nine cases with previously established diagnoses of feline VPs, BISC, invasive SCC and AK were selected from a dermatopathological database. A critical re-evaluation of diagnosis was performed by defining clear criteria toward carcinomatous vs non-carcinomatous, in situ vs invasive (if carcinomatous) and viral vs actinic. Cases were evaluated for p16 immunolocalisation. The presence of the target viral genes for Felis catus papillomavirus (FcaPV)-1, FcaPV-2, FcaPV-3 and FcaPV-4 was determined by qPCR. The data generated ΔΔCq values, which represent a normalised measure of DNA viral quantity. Samples with a positive ΔΔCq value were submitted to sequence analysis. Results Four VPs, 19 BISCs, four SCCs and one case of AK were included. By ΔΔCq analysis we found that all VPs were positive for FcaPV-1 or FcaPV-2; eight BISCs were positive for FcaPV-1, FcaPV-2 and FcaPV-4. FcaPV-2 was the most prevalent among the group of VPs and BISCs. Conclusions and relevance Using the ΔΔCq method we report the first evidence of FcaPV-1, FcaPV-2 and FcaPV-4 in Italy. FcaPV-2 was the most frequently detected; to a lesser extent, FcaPV-1 and FcaPV-4 were detected in the examined samples. FcaPV-3 was never associated with viral-induced lesions by ΔΔCq investigation. Compared with conventional PCR the ΔΔCq method has the advantage of establishing a possible role of the virus in the outcome of infection.
目的 本研究旨在通过定量聚合酶链反应(qPCR)调查猫病毒性斑块(VPs)、鲍温样原位癌(BISC)、鳞状细胞癌(SCC)和光化性角化病(AK)中乳头瘤病毒的存在情况。方法 从皮肤病理学数据库中选取29例先前已确诊为猫VPs、BISC、浸润性SCC和AK的病例。通过定义癌性与非癌性、原位与浸润性(如果是癌性)以及病毒性与光化性的明确标准,对诊断进行了严格的重新评估。对病例进行p16免疫定位评估。通过qPCR确定猫乳头瘤病毒(FcaPV)-1、FcaPV-2、FcaPV-3和FcaPV-4的靶病毒基因的存在情况。所生成的数据为ΔΔCq值,它代表DNA病毒量的标准化测量值。ΔΔCq值为阳性的样本进行序列分析。结果 纳入了4例VPs、19例BISCs、4例SCCs和1例AK。通过ΔΔCq分析,我们发现所有VPs对FcaPV-1或FcaPV-2呈阳性;8例BISCs对FcaPV-1、FcaPV-2和FcaPV-4呈阳性。FcaPV-2在VPs和BISCs组中最为普遍。结论及意义 使用ΔΔCq方法,我们报告了意大利FcaPV-1、FcaPV-2和FcaPV-4的首个证据。FcaPV-2检测频率最高;在所检测的样本中,FcaPV-1和FcaPV-4的检测频率较低。通过ΔΔCq研究,FcaPV-3从未与病毒诱导的病变相关联。与传统PCR相比,ΔΔCq方法的优势在于能够确定病毒在感染结果中的可能作用。