摘要
目的探讨应用HIV-1核酸定量和套式RT-PCR检测技术,对未随访到第2份血样的成年HIV-1抗体阳性不确定样本的检测可行性,及其在艾滋病感染实验室诊断中的作用。方法选取2016年1月—2018年7月经确证实验室Western Blot(WB)法检测为HIV-1不确定样本,2~4周后随访但未获得第2份血浆,无法进行进一步确证实验的11份成年人血浆样品,采用荧光定量PCR法和RT-PCR法进行检测,并计算直接诊断阳性率,与WB法结果进行比较。结果11份未随访到第2份血样的HIV-1成年抗体阳性不确定样本,经WB法确证均为阴性;荧光定量PCR法直接诊断阳性率为63.6%(7/11),CT值为18.8~40.9,病毒载量为5.14×10^7~26.73IU/mL,其中6份>5000IU/mL,占54.5%。对7份荧光定量PCR阳性样本进行RT-PCR扩增,直接诊断阳性率为54.5%(6/11);6份病毒载量>5000IU/mL的标本均获得特异扩增片段,1份病毒载量为26.73IU/mL样品未获得RT-PCR扩增片段。荧光定量PCR法、RT-PCR法直接诊断率均远高于WB(0),且第1份血样就可以做出诊断。结论荧光定量PCR和RT-PCR法检测HIV-1核酸敏感度高于传统WB法,可应用于无法获得第2份随访血样的HIV-1不确定者,尤其是病毒载量>5000IU/mL的标本的检测。
Objective To explore the feasibility of using HIV-1 nucleic acid quantitative detection and nested RT-PCR detection technology to analyze HIV-1 antibody uncertain adults failed to obtain a second plasma and its role in laboratory diagnosis of HIV infection.Methods From Jan 2016 to Jul 2018,the Western blot(WB)analysis was used to detect the HIV-1 antibody uncertain samples in the confirmation laboratory.After 2-4 weeks of follow-up,11 adults failed to obtain the second plasma samples for further confirmation.The fluorescent quantitative PCR and RT-PCR methods were used for analysis;the positive rate of direct diagnosis was calculated and compared with the results of WB method.Results The 11 uncertain samples showed negative results by WB analysis.The positive rate of direct diagnosis was 63.6%(7/11),the CT values were 18.8-40.9,the viral loads were 5.14×10^7-26.73 IU/mL,of which 6 were more than 5000 IU/mL,accounting for 54.5%.RTPCR amplification was carried out on 7 fluorescent quantitative PCR positive samples,the positive rate of direct diagnosis was 54.5%(6/11);specific amplification fragments were obtained from 6 samples with viral load>5000 IU/mL,and no RTPCR amplification fragment was obtained from 1 sample with viral load of 26.73 IU/mL.The direct diagnosis rates of fluorescence quantitative PCR and RT-PCR were much higher than WB(0),the first blood sample can deliver the diagnosis results.Conclusion The sensitivity of fluoresence quantitative PCR and RT-PCR to detect HIV-1 nucleic acid is higher than traditional WB method.It can be used for diagnosis of HIV-1 uncertain cases who failed to obtain the second follow-up blood sample,especially for the samples with viral load>5000 IU/mL.
作者
葛藤
许文炯
董潇潇
王燕
董晓庆
季建强
张洪英
GE Teng;XU Wen-jiong;EXDNG Xiao-xiao;WANG Yan;DONG Xiao-qing;JI Jian-qiang;ZHANG Hong-ying(Nanjing Municipal Center for Disease Control and Prevention,Jiangsu Nanjing 210003,China)
出处
《江苏预防医学》
CAS
2019年第6期639-641,共3页
Jiangsu Journal of Preventive Medicine
基金
江苏省卫计委预防医学课题(Y2015002)
关键词
人类免疫缺陷病毒
核酸检测
WB
PCR
病毒载量
HIV-1不确定
Human immunodeficiency virus
Nucleic acid detection
Western Blot
PCR
Viral load
HIV-1 antibody uncertain patients
