新疆伊宁市912例HIV-1型病毒载量定量检测结果分析

目的分析新疆伊宁市人民医院抗病毒中心912例HIV感染者HIV-1型病毒载量定量检测结果,了解HIV感染者体内病毒载量、病毒载量与临床指征之间的相互关系,为艾滋病的预防和抗病毒治疗提供基础数据。方法采用分枝DNA信号放大系统(bDNA)技术,利用SIEMENS公司VERSANT-440分子系统,2012年7月～2013年2月检测伊宁市912例HIV感染者HIV-1型病毒载量拷贝数。结果伊宁市912例HIV感染者HIV-1病毒载量分布状况:<500 c/ml 661例(72.4%),501～3 000 c/ml 48例(5.3%),3 001～10 000 c/ml 82例(9.0%),10 001～30 000 c/ml 49例(5.4%),30 000～3 5000 c/ml 6例(0.7%),>35 000 c/ml 66例(7.2%)。结论 HIV-1型病毒载量定量检测是判定HIV感染者疾病进程,评估临床抗病毒治疗的重要指标。     

血样的不同处理对HIV-1病毒载量检测值的影响

目的观察血样本存放时间、存放温度及冻融次数对HIV病毒载量检测值的影响。方法收集同一HIV感染者的抗凝血标本,在不同时间分离血浆,并在不同温度下保存不同时间或经过不同次数的冻融,应用HIV-1核酸序列扩增技术(NASBA法)检测各样本的病毒载量进行比较。结果与全血样本相比,以血浆形式保存的样本HIV病毒载量更加稳定;病毒载量的下降主要发生在采血后的6h内;-20℃保存的血浆样本反复冻融3次,病毒载量会显著下降。结论对于运送HIV病人的外周血进行病毒载量检测时,应尽量以低温和血浆形式保存,并尽快的送到检测单位。     

HSV-2治疗可降低HIV-1病毒载量

据Medscape．com2月21日报道（原载N Engl J Med2007；356：790—799，854—856），研究发现，双重感染HIV—1和疱疹病毒-2（HSV-2）的女性患者，在应用伐昔洛韦抗HSV治疗后可降低血液和生殖器分泌物中HIV-1的RNA水平。     

保存时间对血浆样本HIV-1病毒载量测定值的影响研究

目的观察保存时间对血浆样本艾滋病病毒Ⅰ型（HIV-1）病毒载量测定值的影响。方法应用HIV-1核酸序列扩增系统技术（NASBA）对87份-20℃保存的HIV-1抗体阳性血浆样本分组，分别在第0、3、6、11、12、13、14个月进行HIV-1病毒载量检测。结果-20℃保存3个月的样本病毒载量无明显降低，平均下降0．210Log值（P〉0．05），6个月以后的样本病毒载量明显降低，平均下降0．256Log值（P〈0．05），11、12、13个月以后的样本病毒载量明显降低，平均下降大于0．428Log值（P〈0．05），但病毒载量下降幅度与原始病毒载量无关（P〉0．05）。结论血浆样本在-20℃条件下保存6个月以上HIV-1RNA病毒载量水平降低明显，已不能真实反映原始样本病毒载量水平，用于病毒载量检测的血浆样本在-20℃冰柜中不宜超过3个月。     

Xpert检测HIV-1病毒载量假阴性1例

<正>HIV-1病毒载量测定对预测疾病进程、评估治疗效果、指导治疗方案调整有着重要的意义,也可作为HIV感染诊断的补充试验,用于急性期/窗口期诊断、晚期患者诊断、HIV感染诊断和小于18月龄的婴幼儿HIV感染诊断^[1]。实时荧光定量PCR扩增技术是目前临床广泛使用的HIV-1病毒载量检测技术之一,     

HIV-1病毒载量突然升高可能预示患者存在重复感染

研究人员在近期出版的《获得性免疫缺陷综合征》上发表文章指出，当那些未接受治疗的患者突然出现血浆HIV-1病毒载量的迅速升高，就有可能预示着患者存在HIV-1的重复感染，尽管这种情况非常罕见。     

不同HIV-1病毒载量定量检测方法的比较研究

目的 评价不同方法检测Ⅰ型艾滋病病毒(HIV-1)病毒载量的相关性及一致性,探讨国产实时荧光核酸定量检测试剂盒用于临床检测HIV-1病毒载量的可行性.方法 收集61份未治疗的HIV感染者/艾滋病(AIDS)病人血浆标本及57份抗病毒治疗后的血浆标本,使用深圳匹基生物工程股份有限公司实时荧光聚合酶链反应(实时荧光-PCR...     

影响血样样本中HIV-1病毒载量检测值的因素分析

随着我国艾滋病(AIDS)病人治疗的人数逐年增加,全国艾滋病病毒(HIV)病毒载量检测需求迅速上升,HIV-1病毒载量的检测仪将在全国各省确认中心实验室和艾滋病定点医院得到广泛使用。病毒载量检测值的准确性受样品采集、分离、保存和检测方法等多种因素的影响,直接影响病人治疗效果的评价和监测分析感染阶段。该文综述了国内外有关从样本采集到保存的不同环节,对HIVRNA检测值造成影响的相关因素研究结果,以兹为相关临床检测或研究提供参考。     

bDNA和NASBA法定量检测HIV-1病毒载量的比较研究

目的比较分支DNA杂交实验(branched DNA,bDNA)和核酸序列扩增实验(Nucleic acid sequence-based am-plification,NASBA)两种方法检测人类免疫缺陷病毒1型病毒(HIV-1)病毒载量间的一致性。方法对25例HIV感染者/艾滋病(Acquired immune deficiency syndrome,AIDS)患者血浆标本同时用bDNA法和NASBA法检测HIV-1病毒载量,并用流式细胞术检测患者外周血CD4+、CD8+T淋巴细胞。结果bDNA法及NASBA法测得HIV-1病毒载量平均值分别为(4.398±0.580)log拷贝数/ml和(4.488±0.602)log拷贝数/ml,差异无统计学意义(t=1.210,P>0.05);两种方法检测出的病毒载量呈显著直线相关(r=0.8004,P<0.001),且与患者的CD4+T细胞数及CD4+/CD8+比值均呈显著直线负相关。结论bDNA法和NASBA法检测HIV-1病毒载量具有高度一致性,在实际工作中均可选用。     

滤纸干血片提取并检测HIV-1DNA的研究进展

人类免疫缺陷病毒（Human immunodeficiency virus,HIV）是艾滋病（AIDS）的病原体.HIV属于逆转录病毒,进入人体后的主要攻击对象是带CD4受体的T淋巴细胞和巨噬细胞,进入细胞后逆转录成cDNA,再与人的DNA整合在一起成为前病毒DNA,最终由于免疫功能遭到严重破坏,发生各种机会性感染或肿瘤而死亡,而且其病死率极高.截止2004年底,全球有2 610万人死于AIDS,故AIDS有“21世纪瘟疫”之称.     

HIV-1 viral diversity and its implications for viral load testing: review of current platforms

The 2008 Recommendations for care of the International AIDS Society reaffirmed the importance of both accurate and sensitive viral load assessment, and by necessity, access to viral load assays. HIV-1 viral load testing is considered essential when initiating antiretroviral therapy (ART), when monitoring ART response, and when considering switching ART regimens. The demand for accurate, reproducible, and cost-effective viral load assays is therefore a global issue. Although the North American and Western European experience has historically been with HIV-1 group M subtype B virus, this paradigm is changing rapidly as migrants and refugees from developing countries with non-B subtype infections often now present for care in the developed world, and travelers to developing countries acquire non-B subtype infection abroad and present for care at home. Awareness of any clinical or laboratory differences between the common HIV-1 group M subtype B and the newer HIV-1 strains being seen in practice is therefore increasingly important. This review of current HIV-1 viral load testing is focused on the potential value of a standardized genotype assignment for HIV-1 viral subtypes, regular monitoring of the performance of available commercial HIV viral load assays on emerging non-B HIV subtypes, circulating recombinant forms (CRFs) and unique recombinant forms (URFs), and a discussion of the implications for resource-limited settings.     

两种HIV-1核酸定量检测试剂盒v2．0的对比研究

目的 比较罗氏COMBAS AmpliPrep／COMBAS TaqMan HIV—1 Test version 2．0（简称TagMan v2．0试剂）和生物梅里埃NucliSENS EasyQ HIV-1 v2．0（简称EasyQ v2．0试剂）两种试剂,检测艾滋病病毒Ⅰ型（HIV-1）病毒载量间的相关性和一致性。方法 对40份血浆样本采用TaqMan EasyQ v2．0试剂和EasyQ v2．0试剂分别检测HIV-1病毒载量。统计学处理采用配对t检验、回归分析和Bland—Altman分析。结果TaqMan v2．0和EasyQ v2．0试剂测得的病毒载量均值分别为（4．41±0．72）log10拷贝／mL和（3．75±0．75）log10拷贝／mL,差异有统计学意义（t=10．441,P〈0．001）。对两种试剂的检测结果进行回归分析表明,两种试剂有较强的相关性（R2=0．817）。用Bland—Altman分析比较两种试剂检测结果的差异均值,结果具有较好的一致性。结论TaqMan v2．0和EasyQ v2．0两种试剂盒在检测HIV-1病毒载量时具有较好的相关性和一致性。     

Analytical and clinical sensitivity of the Procleix Ultrio HIV-1/HCV/HBV assay in samples with a low viral load

BACKGROUND AND OBJECTIVES: The Procleix Ultrio human immunodeficiency virus type 1 (HIV-1)/hepatitis C virus (HCV)/hepatitis B virus (HBV) (Ultrio) assay simultaneously detects HIV-1 RNA, HCV RNA and HBV DNA in individual blood donations. The main objective of the study was to assess the analytical and clinical sensitivity of the multiplex and discriminatory probe assays in samples with a low viral load. MATERIAL AND METHODS: The VQC HIV RNA genotype B, HCV RNA genotype 1 and HBV DNA genotype A standard dilutions were tested in 26 repeats. The probability of detection by Ultrio was compared with previously obtained data of the Procleix Duplex HIV-1/HCV assay on the same reference panels. A selection of 121 anti-HIV-1, 138 anti-HCV and 190 HBsAg positive samples from patients receiving antiviral therapy were tested. The majority of patient samples had a viral load below the detection limit of the diagnostic nucleic acid test assays, which made them suitable to evaluate the performance of the multiplex and discriminatory assays on yield cases with a similar low viral load. RESULTS: The 95% and 50% detection end-points of the Ultrio assay along with the corresponding 95% confidence intervals are 53.7 (32.9-117.2) and 8.6 (6.2-12.1) geq/ml for HIV-1 RNA, 30.3 (19.0-62.4) and 5.2 (3.7-7.2) geq/ml for HCV RNA and 393.7 (147.9-6978) and 54.5 (22.4-143.8) geq/ml for HBV DNA. The analytical sensitivity of Ultrio expressed as a potency factor relative to previously obtained Duplex results on the same HIV-1 RNA and HCV-RNA standard dilutions was 1.09 (0.20-6.10) and 1.11 (0.21-5.89), respectively. The assay detected all 22 HIV-1 infected patients with viral load > 50 copies/ml, and 41 of 99 patients (41%) with viral load < 50 copies/ml, of which 23 (56%) were detected by the discriminatory assay. All 47 patients with HCV RNA load > 521 IU/ml and 10/91 polymerase chain reaction-negative patients with viral load < 50 IU/ml tested positive in Ultrio assay of which five were missed in the discriminatory test. The assay detected 53/55 HBV infected patients (96%) with viral load > 250 copies/ml and 108/135 patients (80%) with viral load < 250 copies/ml of which 17 (16%) were missed by the discriminatory test. CONCLUSIONS: The new Procleix Ultrio assay is as sensitive as the Procleix Duplex assay for HIV-1 and HCV detection meeting the requirements of universal guidelines. The ability of the assay to detect HBV DNA in low viral load samples could be useful for screening blood. Inevitable negative results of discriminatory probe assays caused by stochastic sample variation will reduce the chance of recognizing low viraemic blood donors detected by individual donation nucleic acid test.     

A case series of third-trimester raltegravir initiation: Impact on maternal HIV-1 viral load and obstetrical outcomes

OBJECTIVE:

To describe the impact of initiating raltegravir (RAL)-containing combination antiretroviral therapy (cART) regimens on HIV viral load (VL) in pregnant women who have high or suboptimal VL suppression late in pregnancy.

METHODS:

HIV-infected pregnant women who started RAL-containing cART after 28 weeks’ gestation from 2007 to 2013 were identified in two university hospital centres.

RESULTS AND DISCUSSION:

Eleven HIV-infected women started RAL at a median gestational age of 35.7 weeks (range 31.1 to 38.0 weeks). Indications for RAL initiation were late presentation in pregnancy (n=4) and suboptimal VL suppression secondary to poor adherence or viral resistance (n=7). Mean VL at the time of RAL initiation was 73,959 copies/mL (range <40 to 523,975 copies/mL). Patients received RAL for a median of 20 days (range one to 71 days). The mean decline in VL from the time of RAL initiation to delivery was 1.93 log, excluding one patient who received only one RAL dose and one patient with undetectable VL at the time of RAL initiation. After eight days on RAL, 50% of the women achieved a VL <1000 copies/mL (the threshold for recommended Caesarean section to reduce the risk for perinatal transmission). There were no cases of perinatal HIV transmission.

CONCLUSION:

The present study provides preliminary data to support the use of RAL-containing cART to expedite HIV-1 VL reduction in women who have a high VL or suboptimal VL suppression late in pregnancy, and to decrease the risk of HIV perinatal transmission while avoiding Caesarean section. Further assessment of RAL safety during pregnancy is warranted.     

双杂交技术在HIV-1研究中的应用进展

随着蛋白组学和基因组学研究的进一步深化,双杂交技术也迅猛发展,在研究细胞内蛋白质分子之间或病毒-宿主之间的相互作用等方面发挥重要作用。本文简述双杂交技术的基本原理、主要类型及其在逆转录病毒HIV-1研究中的应用进展。     

Proviral HIV-1 dynamics and evolution in patients receiving efficient long-term antiretroviral combination therapy

Different experimental approaches have shown that, despite plasma viral loads under the threshold of detection, HIV-1 frequently continues to replicate in patients receiving potent antiretroviral therapy. However, whether this low-grade viral replication is sufficient for the generation of new major quasispecies has not been studied. Thus, in order to evaluate the extent of variation in the major proviral HIV-1 population, we monitored proviral DNA sequences in such patients over a time period of up to 30 months.
Methods   DNA was extracted from peripheral blood mononuclear cells (PBMC) and the V3 region was amplified by nested polymerase chain reaction (PCR) and directly sequenced. Additionally, both HIV-1 RNA and DNA levels and CD4⁺ T-lymphocyte counts were monitored.
Results   Analysing the V3 gene sequences of 17 patients, we observed a sequence evolution in nine patients. Interestingly, the majority of these changes (77%) occurred in the first interval following the initiation of therapy and despite signs of ongoing replication the proviral DNA levels continued to decrease in all patients.
Conclusions   Our data suggest that, although available data report that HIV-1 continues to replicate in patients with undetectable viraemia, the extent of viral replication in many of these patients is not sufficient to result in changes in the major viral population.     

The effects of an HIV-1 immunogen (Remune) on viral load, CD4 cell counts and HIV-specific immunity in a double-blind, randomized, adjuvant-controlled subset study in HIV infected subjects regardless of concomitant antiviral drugs

Objective   We examined the activity of an HIV-1 immunogen (Remune) on viral load, CD4 cells and HIV-1 specific immunity.
Methods   Plasma and peripheral blood mononuclear cells were obtained in a predefined random subset of subjects ( n  = 252) from a multicentre, double-blind, adjuvant-controlled phase III clinical endpoint study.
Results   The subjects treated with the HIV-1 immunogen had a significantly greater decline in viral load at multiple time points ( P  < 0.05), a trend towards increased CD4+ T cell counts and significantly enhanced HIV-1 specific immune responses as measured by HIV-1 lymphocyte proliferation ( P  < 0.001) compared to the adjuvant control group. Furthermore, in the HIV-1 immunogen treated group, enhanced HIV-1 specific lymphocyte proliferative immune responses were associated with decreased HIV-1 plasma RNA.
Conclusion   These results suggest that, in a predefined, random subset of subjects, a beneficial effect of the HIV-1 immunogen was observed on viral load, CD4+ T cells, and HIV-specific immunity. These differences were observed in a background of multiple drug therapies. Ongoing trials are evaluating the effect of the combination of this HIV-1 specific, immune-based therapy with potent antiviral drug therapy on virological outcomes.