Localization of Fibropapilloma-associated Turtle Herpesvirus in Green Turtles (Chelonia mydas) by In-Situ Hybridization
Localization of Fibropapilloma-associated Turtle Herpesvirus in Green Turtles (Chelonia mydas) by In-Situ Hybridization
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DOI：
10.1016/j.jcpa.2008.07.003
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发表时间：
2008-11-01
影响因子：
0.8
通讯作者：
Brown, C. C.
中科院分区：
文献类型：
--
作者：
Kang, K. I.;Torres-Velez, F. J.;Brown, C. C.
纤维乳头瘤相关海龟疱疹病毒（FPTHV）是海龟纤维乳头瘤病（FP）的假定病原体。已分别通过聚合酶链反应（PCR）和逆转录酶-PCR（RT-PCR）检测到病毒的病灶内DNA和RNA，但尚未解决病毒在肿瘤内的确切位置和分布。本研究利用原位杂交技术研究了FPTHV的转录活性和定位。从波多黎各岛（库莱布拉和库莱布里塔）检查的105只绿色海龟（Chelonia mydas）的皮肤或结膜中获得25个肿瘤。这些病变包括19个纤维乳头状瘤和6个纤维瘤。FPTHV mRNA转录检测ISH在三个纤维乳头状瘤，阳性反应仅限于上皮细胞簇的细胞核。核糖核酸探针原位杂交结合变性法在14个肿瘤中检测到病毒DNA，包括纤维乳头状瘤和纤维瘤。信号仅限于棘层上皮细胞的细胞核，在肿瘤的上皮下纤维区未观察到。这些结果表明，FPTHV存在于上皮细胞和转录活跃的纤维乳头状瘤。(c)2008爱思唯尔有限公司保留所有权利。
Fibropapilloma-associated turtle herpesvirus (FPTHV) is the presumed aetiological agent of sea turtle fibro-papillomatosis (FP). Intralesional DNA and RNA of the virus have been detected by polymerase chain reaction (PCR) and reverse transcriptase-PCR (RT-PCR), respectively, but the exact location and distribution of the virus within the tumours have not been addressed. In this study, in-situ hybridization (ISH) was used to investigate viral transcriptional activity and localization of FPTHV. Twenty-five tumours were obtained from the skin or conjunctiva of 105 green turtles (Chelonia mydas) examined on two islands in Puerto Rico (Culebra and Culebrita). These lesions comprised 19 fibropapillomas and six fibromas. FPTHV mRNA transcripts were detected by ISH in three fibropapillomas, with positive reactions confined to the nuclei of clusters of epithelial cells. Viral DNA was detected by riboprobe ISH combined with denaturation in 14 tumours, including both fibropapillomas and fibromas. Signals were confined to the nuclei of acanthotic epithelial cells and were not seen in the subepithelial fibrous areas of the tumours. These results suggest that FPTHV is present in epithelial cells and transcriptionally active in fibropapillomas. (c) 2008 Elsevier Ltd. All rights reserved.