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利用实时荧光定量PCR对海龟正常组织和纤维乳头瘤中的疱疹病毒序列进行定量分析。

Quantitative analysis of herpesvirus sequences from normal tissue and fibropapillomas of marine turtles with real-time PCR.

作者信息

Quackenbush S L, Casey R N, Murcek R J, Paul T A, Work T M, Limpus C J, Chaves A, duToit L, Perez J V, Aguirre A A, Spraker T R, Horrocks J A, Vermeer L A, Balazs G H, Casey J W

机构信息

Department of Molecular Biosciences, University of Kansas, Lawrence, Kansas 66045-2106, USA.

出版信息

Virology. 2001 Aug 15;287(1):105-11. doi: 10.1006/viro.2001.1023.

Abstract

Quantitative real-time PCR has been used to measure fibropapilloma-associated turtle herpesvirus (FPTHV) pol DNA loads in fibropapillomas, fibromas, and uninvolved tissues of green, loggerhead, and olive ridley turtles from Hawaii, Florida, Costa Rica, Australia, Mexico, and the West Indies. The viral DNA loads from tumors obtained from terminal animals were relatively homogeneous (range 2-20 copies/cell), whereas DNA copy numbers from biopsied tumors and skin of otherwise healthy turtles displayed a wide variation (range 0.001-170 copies/cell) and may reflect the stage of tumor development. FPTHV DNA loads in tumors were 2.5-4.5 logs higher than in uninvolved skin from the same animal regardless of geographic location, further implying a role for FPTHV in the etiology of fibropapillomatosis. Although FPTHV pol sequences amplified from tumors are highly related to each other, single signature amino acid substitutions distinguish the Australia/Hawaii, Mexico/Costa Rica, and Florida/Caribbean groups.

摘要

定量实时聚合酶链反应已被用于测量来自夏威夷、佛罗里达、哥斯达黎加、澳大利亚、墨西哥和西印度群岛的绿海龟、蠵龟和榄蠵龟的纤维瘤、纤维乳头状瘤及未受累组织中与纤维乳头状瘤相关的海龟疱疹病毒(FPTHV)聚合酶(pol)基因的DNA载量。来自濒死动物肿瘤的病毒DNA载量相对一致(范围为2 - 20拷贝/细胞),而来自活检肿瘤及健康海龟皮肤的DNA拷贝数则呈现出广泛的变化(范围为0.001 - 170拷贝/细胞),这可能反映了肿瘤发展的阶段。无论地理位置如何,肿瘤中的FPTHV DNA载量都比同一动物未受累皮肤中的高2.5 - 4.5个对数,这进一步表明FPTHV在纤维乳头状瘤病的病因学中起作用。尽管从肿瘤中扩增出的FPTHV pol序列彼此高度相关,但澳大利亚/夏威夷、墨西哥/哥斯达黎加和佛罗里达/加勒比海群体之间存在单个特征性氨基酸替换。

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