不同核酸提取方法检测HIV病毒载量和耐药基因的研究.doc
《不同核酸提取方法检测HIV病毒载量和耐药基因的研究.doc》由会员分享,可在线阅读,更多相关《不同核酸提取方法检测HIV病毒载量和耐药基因的研究.doc(7页珍藏版)》请在三一文库上搜索。
1、不同核酸提取方法检测HIV病毒载量和耐药基因的研究Abstract Objective To compare the efficiency of HIV RNA extracted by five kinds of nucleic acid extraction methods. Methods Blood samples were collected from HIV infected persons in 2015. Used different methods of nucleic acid extraction to detect the Ct value and the load v
2、alue. The detection rate, the sensitivity, the coefficient of variation, the stability, and the drug resistance were detected. Results Domestic magnetic bead method had the highest detection rate in five kinds of nucleic acid method, there was no statistically significant difference of 5 kinds of me
3、thod (2=7.98, P=0.09). Ct values from small to large order: domestic magnetic beadscentrifugal column methodlarge quantities of extraction methodimported magnetic beadsphenol/chloroform cracking. Repeatability tests showed that domestic magnetic beads was stable; Nucleic acid gel showed domestic mag
4、netic beads to extract RNA concentration was highest; Extraction method did not affect the detection of drug resistance. Conclusion Five kinds of nucleic acid extraction methods are able to detect HIV information. The viral nucleic acid extraction using magnetic beads method, sensitivity and stabili
5、ty are higher. Key words Nucleic acid extraction; Quantitative PCR; HIV load; Resistance gene; Magnetic beads; HIV RNA 人类免疫缺陷病毒(Human Immunodeficiency Virus,HIV)已在全球肆虐40余年。据WHO统计,至2015年底预计有1500万感染者接受抗病毒治疗1。根据艾滋病治疗手册,规定接受治疗的艾滋病患者必须定期检测病毒载量,用于反映药物疗效以及预防耐药的发生。抗病毒治疗可以降低艾滋病患者血液中的病毒载量,恢复部分免疫功能。但是长期接受抗病毒治疗
6、、患者依从性差等均易导致对治疗药物产生耐药性,继而加剧耐药株的传播。非洲某些地区新发病者耐药性高达15.2%,诸多一线治疗药物失效2,3。 目前HIV病毒载量检测一般为RNA,克隆病毒基因片段关键在于高效提取RNA核酸。此外,对基因分型溯源分析、CD4+T细胞受体CCR532/32、肿瘤循环DNA等研究也要求完整的序列片段4-6。核酸RNA模板提取的过程是影响后续试验结果的关键,尤其是对病毒水平较低的样本影响会更明显。基层疾控系统常用的病毒核酸提取方法主要有磁珠法和离心柱法7,8,多为进口试剂,进口试剂价格昂贵,近年来,国内基于实时荧光定量技术与不同的核酸提取方法进行研制,凯杰、达安基因、珠海
- 配套讲稿:
如PPT文件的首页显示word图标,表示该PPT已包含配套word讲稿。双击word图标可打开word文档。
- 特殊限制:
部分文档作品中含有的国旗、国徽等图片,仅作为作品整体效果示例展示,禁止商用。设计者仅对作品中独创性部分享有著作权。
- 关 键 词:
- 不同 核酸 提取 方法 检测 HIV 病毒 耐药 基因 研究
链接地址:https://www.31doc.com/p-1593022.html