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【资讯翻译】Development and application of a rapid detection system

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发表于 2016-6-13 11:35:24 | 显示全部楼层 |阅读模式
Development and application of a rapid detection system for human papillomavirus and Herpes simplex virus -2 by loop-mediated isothermal amplification assay.
Human papillomavirus (HPV) infection is an important factor that causes cervical cancer and non-melanoma skin cancer (NMSC), while HSV-2 plays an important role when HR-HPV triggers the cancer. Thus, a quick and convenient assay in the detection of HPV and HSV-2in the screening of HPV and HSV-2 infection is required. Two respective HPV and HSV-2 detection methods were established based on loop-mediated isothermal amplification (LAMP) assay. Specific outer primers, inner primers, and loop primers were designed according to the conserved domains of HPV and HSV-2 genomes, respectively, while degenerate primers were used for HPV assay. After optimizing the reaction conditions, the results were observed by LAMP Tubidimeter real-time LA-320. Standard plasmids HPV-L-P and HSV-2-L-P were cloned and used in sensitivity tests of HPV LAMP and HSV-2 LAMP, respectively. Fifty samples of actinic keratosis (AK), 20 samples of squamous cell carcinoma (SCC), 50 samples of basal cell carcinoma (BCC) and 20 samples of seborrheic keratosis (SK) were detected by HPV assay. Seventy three clinical samples of vaginitis, chronic cervicitis, cervical intraepithelial neoplasias and cervical cancer level positive were detected with HPV and HSV-2 assays. The reaction conditions of two assays were the same with a reaction temperature of 63 °C and a reaction time of 45 min. The sensitivity of HPV LAMP assay was 10 copies/μL, while that of the HSV-2 LAMP assay was 100 copies/μL. No cross-reactivity was observed. The HPV positive rates of AK, SCC, BCC and SK samples were 80% (40/50), 75% (15/20), 44% (22/50) and 21% (15/72), respectively. As an economic and quick diagnostic tool, LAMP assay is conducive to the extensive screening of HPV and HSV-2 and has huge potential to be promoted in resource-limited hospitals.


http://www.ncbi.nlm.nih.gov/pubmed/27287497

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发表于 2016-6-13 12:00:59 | 显示全部楼层
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发表于 2016-6-13 15:38:13 | 显示全部楼层
利用环介导等温扩增实验针对人乳头瘤病毒和单纯疱疹病毒-2的快速检测系统的开发与应用
人乳头瘤病毒(HPV)感染是是引起宫颈癌和非黑素瘤皮肤癌(NMSC)的一个重要的因素,HSV-2在HR-HPV触发癌症时起着重要的作用。因此,一个快速和方便的检测实验在检测HPV和hsv-2在 HPV和HSV-2感染的筛查是必要的。两种HPV和HSV-2检测方法建立基于环介导等温扩增(LAMP)检测。特异外引物,内引物,环引物按照HPV和HSV-2病毒基因组保守结构域设计,此外简并引物可进行HPV检测。优化反应条件后,通过LAMP Tubidimeter real-time LA-320可观察到结果。标准品HPV-L-P和HSV-2-L-P克隆分别用于 HPV LAMP和 HSV-2 LAMP敏感性试验。50例光线性角化病(AK),20份鳞状细胞癌(SCC)样本,50份基底细胞癌(BCC)样品和20份脂溢性角化病的样品(SK)进行HPV检测。73例阴道炎、慢性宫颈炎、宫颈上皮内瘤变和宫颈癌通过HPV和HSV-2检测被发现。两次实验反应条件的反应温度63°C和反应时间45分钟完全一致。HPV LAMP检测的灵敏度为10 copies/μL,而HSV-2 LAMP检测100 copies/μL。无交叉反应现象产生。AK,SCC, BCC和SK样品中HPV阳性分别为80%(40/50)、75%(15/20)、44%(22/50)和21%(15/72)。作为一个经济和快速的诊断工具,LAMP方法有利于HPV和HSV-2的广泛筛选,在资源有限的医院推广潜力巨大。

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 楼主| 发表于 2016-6-14 09:50:06 | 显示全部楼层
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