Development of a broad spectrum PCR assay for papillomaviruses and its application in screening lung cancer biopsies.
Development of a broad spectrum PCR assay for papillomaviruses and its application in screening lung cancer biopsies.
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乳头瘤病毒广谱 PCR 检测的开发及其在筛查肺癌活检中的应用。
DOI：
10.1099/0022-1317-75-5-1149
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发表时间：
1994
期刊：
影响因子：
--
通讯作者：
de Villiers
中科院分区：
文献类型：
--
作者：
Vladimir Shamanin;Hajo Delius;Ethel;de Villiers
开发了一种 PCR 检测方法来检测已知和尚未鉴定的乳头状瘤病毒 (PV)。为此，我们分析了 45 个人类和 9 个动物 PV 的 L1 和 E1 开放阅读框中的保守氨基酸序列。用于设计引物的候选区域被鉴定为在不同PV中具有最少数量的氨基酸和核苷酸序列变体的区域。 L1 ORF 中的这些区域之前已进行过描述。我们修改了反向和正向引物的序列，以及用作简并探针的寡核苷酸的序列，以覆盖更广泛的 PV 谱。在与 32P 标记的简并寡核苷酸探针杂交后，对人和动物 PV 进行检测的灵敏度是，对于整合的 PV DNA，每个细胞有 1 个基因组拷贝；对于质粒 PV DNA，每个细胞有 10 个基因组拷贝。唯一的例外是人乳头瘤病毒 (HPV) 4 型、HPV60 和 HPV65，其敏感性较低。该组只能通过使用额外的引物来检测。该测定用于分析代表不同组织学类型的 85 个肺癌活检组织。使用该系统，与人胎盘 DNA（阴性对照）和 PV DNA 阳性标准相比，在活检中未检测到 PV DNA 序列。
A PCR assay was developed to detect known and as yet unidentified papillomaviruses (PVs). For this purpose we analysed the conserved amino acid sequences in the L1 and E1 open reading frames of 45 human and nine animal PVs. Candidate regions for the design of a primer were identified as those having the least number of amino acid and nucleotide sequence variants among the different PVs. These regions in the L1 ORF have been described previously. We modified the sequences of the backward and the forward primers, as well as the sequence of the oligonucleotide used as the degenerate probe, in order to cover a broader spectrum of PVs. The sensitivity of the assay for the human and animal PVs tested after hybridization with a 32P-labelled degenerate oligonucleotide probe was one genome copy per cell for integrated PV DNA and 10 genome copies per cell for plasmid PV DNA. The only exceptions were human papillomavirus (HPV) type 4, HPV60 and HPV65, for which a lower sensitivity was obtained. This group could be detected only by using additional primers. The assay was used to analyse 85 lung cancer biopsies representing different histological types. Using this system no PV DNA sequences were detected in the biopsies when compared with human placental DNA (a negative control) and PV DNA-positive standards.
复发性喉乳头状瘤病患者肺癌中的人乳头瘤病毒 6a 型 DNA 具有部分重复的特征。
DOI：
10.1099/0022-1317-73-2-423
发表时间：
1992
期刊：
The Journal of general virology
影响因子：
--
作者：
DiLorenzo,TP;Tamsen,A;Abramson,AL;Steinberg,BM
通讯作者：
Steinberg,BM