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[使用地高辛标记的DNA探针检测鼻喉乳头瘤中的人乳头瘤病毒基因组]

[Detection of human papillomavirus genome in nasolaryngeal papillomas using digoxigenin labeled DNA probes].

作者信息

Furuta Y, Inuyama Y, Nagashima K

出版信息

Nihon Jibiinkoka Gakkai Kaiho. 1989 Dec;92(12):2055-63. doi: 10.3950/jibiinkoka.92.2055.

Abstract

It is being reported that human papillomavirus (HPV) has been implicated in the pathogenesis of various neoplastic lesions of the genital organs. To investigate the etiological role of HPV and its types in nasolaryngeal papillomas, we retrospectively analyzed HPV genomes by nucleic acid hybridization methods; for detecting DNA and mRNA, we employed the recently developed nonradioactive (digoxigenin labeled) DNA probes and compared the results by radioisotope methods. In total, 43 cases of papillomatous lesions were examined. They were verruca vulgaris of the nasal vestibule (Nr = 2), nasal inverted papilloma (IP, Nr = 26), and laryngeal papilloma (Nr = 15). HPV types examined were type 2, 6, 11, 16 and 18. Two cases of verruca vulgaris were shown to contain HPV-2 DNA and its mRNA by in situ hybridization. HPV-11 DNA was detected in 3 cases (12%) of nasal inverted papilloma whereas HPV-16 was detected in 1 case (4%); the latter case was associated with squamous cell carcinoma. These results suggest that HPV may be implicated in the development of IP, and HPV-16 may play an important role in the malignant transformation of IP. In the cases of multiple laryngeal papilloma (Nr = 8, one juvenile type and 7 adult type), either HPV-6 or HPV-11 was detected at the high rate (6/8, 75%). The presence of the HPV genomes provides strong evidence for the HPV etiology of these laryngeal papillomas. Whereas in the cases of adult single laryngeal papilloma (Nr = 7), HPV was not detected. Technically, the sensitivity of digoxigenin (DIG) labeled DNA probe was almost same as 35S labeled probe by dot blot hybridization, thus we applied DIG labeled probe to Southern blot hybridization with low background. By in situ hybridization using digoxigenin labeled probes, the rates of HPV detection were almost equal to those by 35S labeled probes.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

据报道,人乳头瘤病毒(HPV)与生殖器官的各种肿瘤性病变的发病机制有关。为了研究HPV及其类型在鼻喉乳头状瘤中的病因学作用,我们通过核酸杂交方法对HPV基因组进行了回顾性分析;为了检测DNA和mRNA,我们使用了最近开发的非放射性(地高辛标记)DNA探针,并通过放射性同位素方法比较了结果。总共检查了43例乳头状病变。它们是鼻前庭寻常疣(n = 2)、鼻内翻性乳头状瘤(IP,n = 26)和喉乳头状瘤(n = 15)。检测的HPV类型为2型、6型、11型、16型和18型。通过原位杂交显示,2例寻常疣含有HPV - 2 DNA及其mRNA。在3例(12%)鼻内翻性乳头状瘤中检测到HPV - 11 DNA,而在1例(4%)中检测到HPV - 16;后一例与鳞状细胞癌相关。这些结果表明,HPV可能与IP的发生有关,HPV - 16可能在IP的恶性转化中起重要作用。在多发性喉乳头状瘤病例(n = 8,1例青少年型和7例成人型)中,HPV - 6或HPV - 11的检出率很高(6/8,75%)。HPV基因组的存在为这些喉乳头状瘤的HPV病因提供了有力证据。而在成人单发性喉乳头状瘤病例(n = 7)中,未检测到HPV。从技术上讲,地高辛(DIG)标记的DNA探针通过斑点杂交的灵敏度与35S标记的探针几乎相同,因此我们将DIG标记的探针应用于背景较低的Southern印迹杂交。通过使用地高辛标记的探针进行原位杂交,HPV的检出率几乎与35S标记的探针相同。(摘要截短于250字)

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