刘　敏　陈秀英　梅少林　陈沙彬　雷永良浙江省丽水市疾病预防控制中心，浙江丽水　323000

不同核酸提取方法检测H IV病毒载量和耐药基因的研究

刘敏陈秀英梅少林陈沙彬雷永良▲
浙江省丽水市疾病预防控制中心，浙江丽水323000

目的比较5种核酸提取法提取HIV RNA的效率。方法收集2015年HIV病毒感染者血样，使用不同核酸提取方法平行检测Ct值及载量值，分析检测的检出率、敏感性、变异系数、稳定性，并进行耐药扩增。结果5种提核酸方法检出率国产磁珠法最高，5种方法差异无统计学意义（χ2=7.98，P=0.09）；Ct值从小到大顺序为：国产磁珠法≥离心柱法＞大批量法≥进口磁珠法＞试剂盒自带法；重复性比较表明国产磁珠法检测结果较稳定；核酸胶显示国产磁珠法提取RNA浓度最高；提取方法不影响耐药关键位点检测。 结论5种核酸提取方法均能检测HIV信息，国产磁珠法提取病毒核酸进行检测，敏感性和稳定性均较高。

核酸提取；荧光定量PCR；HIV载量；耐药基因；磁珠；HIV RNA

［Abstract］Objective To compare the efficiency of HIV RNA extracted by five kinds of nucleic acid extraction methods.M ethods Blood sampleswere collected from HIV infected persons in 2015.Used differentmethods of nucleic acid extraction to detect the Ct value and the load value.The detection rate，the sensitivity，the coefficient of variation，the stability，and the drug resistancewere detected.Results Domesticmagnetic bead method had the highest detection rate in five kinds of nucleic acid method，there was no statistically significant difference of 5 kinds ofmethod（χ2=7.98，P= 0.09）.Ct values from small to large order:domestic magnetic beads≥centrifugal column method＞large quantities of extraction method≥imported magnetic beads＞phenol/chloroform cracking.Repeatability tests showed that domestic magnetic beads was stable；Nucleic acid gel showed domestic magnetic 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 usingmagnetic beadsmethod，sensitivity and stability 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、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］。……