核酸定量检测应用于HIV-1感染诊断的研究

目的探讨核酸定量检测在1型艾滋病病毒(HIV-1)感染诊断中的应用。方法选取云南省德宏州216例样本,包括136例HIV-1阳性母亲所生活产婴儿和80例HIV-1阳性成人,分别用HIV-1核酸定性和定量方法进行检测,综合对比分析2种方法的检测结果。结果 216例样本的核酸定性和定量检测试验一致性极好,53例样本(50例成人样本和3例婴儿样本)核酸阳性,血浆病毒载量5 000拷贝/mL。随访50例成人样本的蛋白印迹试验(WB),均为确证HIV-1抗体阳性。163例阴性样本(30例成人样本和133例婴儿样本)核酸阴性,血浆病毒载量低于检测下限。结论当血浆病毒载量5 000拷贝/mL,核酸定量检测试验对于HIV-1感染阳性样本是一种有效的实验室诊断方法。     

美国CDC推荐HIV核酸检测策略与核酸定量检测替代策略

美国疾病控制和预防中心(CDC)2014年发布艾滋病病毒(HIV)核酸检测策略以来,已经在实验室应用,该策略简化了检测流程,使检测结果更加准确。但同时也存在诸多不便于临床推广普及的影响因素。随着核酸检测技术的应用与普及,在2019年艾滋病诊断大会上,美国CDC专家发表了HIV-1核酸定量检测替代策略的研究报告,临床验证了核酸定量检测替代策略的准确性、可行性与快速便利性,提高了HIV核酸定量检测结果在临床工作中的使用价值。     

干血斑HIV-1核酸定性检测的可靠性研究

目的探索干血斑(DBS)核酸定性检测用于HIV-1感染诊断的可靠性。方法同步采集参加自愿咨询检测人员的DBS样本和配套血清样本各230份,DBS样本用于HIV-1核酸定性检测,血清样本进行常规抗体检测,分析比较两种方法的检测结果。结果 DBS初次核酸定性结果与血清初次抗体检测结果一致率为98.70%,Kappa值为0.93,经复核及随访复检后两种方法最终判断结果完全一致,一致率为100%。其中2例抗体不确定样本其核酸定性检测为阳性,随访后WB条带进展,确证为HIV-1抗体阳性;有1例抗体阴性样本其首次核酸定性检测为阳性,双份复核为阴性。结论 DBS核酸定性结果与常规血液抗体检测的感染诊断结果一致性好,且流程简便、准确性高,但需考虑不确定范围。     

HIV抗体检测策略的临床验证结果分析

目的针对《全国艾滋病检测技术规范》(2015年修订版)中的临床诊断的抗体检测策略进行验证,证明"抗体确证试验"作为补充试验时存在局限性,对处于早期感染样本的检测有漏检风险方法对2017年6月至2018年5月期间,本实验室抗体确证试验结果为"HIV抗体阴性"的160份样本,用灵敏度略高的抗体试剂进行复检,按复检结果报告,并对两次检测结果不一致的样本进行核酸检测。结果 160份样本中40份复检为"HIV-1抗体不确定",无"HIV-1抗体阳性",进行核酸检测的样本17/40结果5 000拷贝/mL,2/40结果5 000拷贝/mL,21/40未检出。收集到17份随访复查结果,其中11份"HIV-1抗体阳性",4份"HIV抗体阴性",2份"HIV-1抗体不确定"。结论抗体确证试验检测为"HIV抗体阴性"的样本,不能直接排除HIV感染,建议对"HIV抗体阴性"样本进行区别分析,有必要时进行随访复查或核酸检测。     

HIV-2型感染流行病学诊疗方案及防控策略

艾滋病病毒(HIV)包含HIV-1和HIV-2两种类型。到目前为止,在HIV感染的临床筛查中,发现的感染类型绝大多数为HIV-1型,极少数为HIV-2型。国外HIV-2型主要集中在西非地区,我国偶有发现HIV-2型的感染病例。由于临床中遇到的HIV-2型感染病例数较少,所以对HIV的研究多集中在了HIV-1型上。随着全球化的推进和人员交流日益频繁,HIV-2型感染的风险正在不断增加,临床上遇到的HIV-2型感染病例也会增加,应引起科研人员和临床医生足够的重视。本文从HIV-2型感染的流行病学、诊断方案及抗病毒防治措施等几个方面进行探讨,以期为我国HIV-2型的预防及诊治策略的制定提供参考意见。     

献血者HIV一次筛查与两次筛查对比分析

正《血站技术操作规程(2012版)》献血者血液人类免疫缺陷病毒(HIV)检测采用2个不同生产厂家的酶联免疫法(ELISA)试剂检测HIV-1和HIV-2抗体或联合检测HIV-1和HIV-2抗原和抗体,而新版(2015版)血液检测策略采用2遍血清学方法和1遍核酸检测或1遍血清学方法和1遍核酸检测。多年来我国血站采用2遍ELISA,为此本文分析献血者血液HIV筛查ELISA 1次筛查方法     

HIV-1感染中的白介素-2免疫治疗作用

抗反转录病毒治疗（ART）开始的时机越来越早,但总有免疫重建失败或者Ⅰ型艾滋病病毒（HIV-1）潜伏感染灶无法清除的情况,因此白介素-2（IL-2）目前被广泛研究用于辅助ART治疗、加强免疫重建或者活化潜伏感染的HIV-1。目前比较确定的是：（1）IL-2能延长CD4＋T细胞的半衰期,增加CD4＋T细胞的数量;（2）IL-2合并ART与单独使用ART相比,不能减少HIV-1相关的机会性感染或死亡,临床应用着重于活化HIV-1储存库细胞;（3）IL-2合并ART能有效增加HIV-1储存库细胞的消减速率,外周循环中HIV-1感染的处于静止状态的记忆T细胞数量下降,甚至检测不出来;（4）IL-2的临床应用趋向周期性使用和长期使用。     

提高对HIV-1急性感染和HIV-2诊断能力的检测策略北大核心

针对多年使用的艾滋病病毒(HIV)检测策略存在的问题,以及目前的检测技术研发和应用状况,美国疾病控制和预防中心在2014年6月提出了新的HIV检测策略,联合使用HIV抗原和抗体检测试剂、能够区分HIV-1和HIV-2抗体的诊断试剂以及HIV-1核酸定性诊断试剂,提高对HIV-1急性感染和HIV-2的检出效能,同时减少需要随访的不确定结果,缩短等待时间。文章对这个检测策略进行了全面介绍,并提出制定适合我国的HIV检测策略的建议。     

提高对HIV-1急性感染和HIV-2诊断能力的检测策略

针对多年使用的艾滋病病毒(HIV)检测策略存在的问题,以及目前的检测技术研发和应用状况,美国疾病控制和预防中心在2014年6月提出了新的HIV检测策略,联合使用HIV抗原和抗体检测试剂、能够区分HIV-1和HIV-2抗体的诊断试剂以及HIV-1核酸定性诊断试剂,提高对HIV-1急性感染和HIV-2的检出效能,同时减少需要随访的不确定结果,缩短等待时间。文章对这个检测策略进行了全面介绍,并提出制定适合我国的HIV检测策略的建议。     

核酸定量检测试验应用于HIV-1感染诊断的评价

正1型艾滋病病毒(HIV-1)感染的诊断依赖于实验室检测,不同的检测方法由于标志物的不同而有不同的窗口期。常规的酶联免疫吸附试验(ELISA)+蛋白印迹试验(WB)的检测策略存在早期感染漏检的风险。在可疑人群中应用更高效的检测策略,可以有效地诊断HIV-1的早期感染。对2018年本确证实验室复检的第四代HIV筛查试剂有反应而WB结果为阴性或不确定的样本进行HIV-1核酸定量检     

两代酶联免疫试剂在男男性行为人群HIV检测中的应用

目的通过使用第四代与第三代艾滋病病毒(HIV)抗体酶联免疫试剂检测结果及核酸检测结果的比较,评估其在天津市男性性行为人群(MSM)HIV感染筛查中的应用效果。方法对天津市某区自愿咨询检测点进行咨询检测的男男性行为人群用EDTA抗凝管采集血液样品,同时使用两代HIV抗体筛查试剂进行检测,阳性者进行确证检测,两种筛查试剂检测结果均为阴性的样品,采用集合核酸检测方法进行检测。结果共计检测MSM人群血液样本2 065份,共对87份样品进行了确证试验,1 978份HIV抗体筛查阴性样品经核酸检测,有1份集合样品呈阳性。HIV抗体确证试验为阴性和不确定的6份样品,进行单份核酸检测,有4份HIV-1RNA阳性,病毒载量检测结果均符合HIV急性感染特征。此次研究中的MSM人群HIV感染率为4.16%(86/2 065),95%可信区间3.34%～5.12%。发现5例HIV急性感染者,估计该人群年发病率为3.16%,95%可信区间1.01%～7.35%。使用第三代抗体试剂进行筛查时,总费用为148 360元,其敏感性为94.2%(81/86)、特异性为100%,发现HIV感染者的单位成本为1 832.6元。使用第四代抗体试剂进行筛查时,总费用为166 080元,其敏感性为98.8%(85/86)、特异性为99.9%(1 977/1 979),发现HIV感染者的单位成本为1 953.9元。结论此次研究估测该人群的HIV感染年发病率为1.01%～7.35%,提示其高危性行为程度较高。在该人群中,使用第四代试剂筛查时,HIV感染者发现数比使用第三代试剂增加4.9%,发现HIV感染者的单位成本增加6.2%,提示具有推广应用价值。     

3种新型冠状病毒核酸检测试剂对120份样本检测结果的比较与分析

目的 对市售的3种国产新型冠状病毒核酸检测试剂(实时荧光RT-PCR法)的检测结果进行比较和分析,以供检测机构参考。方法 采用3种核酸检测试剂(实时荧光RT-PCR法)对120份新冠肺炎病人和密切接触者的咽拭子、尿液、唾液和粪便样本进行平行检测,运用SPSS20软件采用卡方检验对检测结果进行分析。结果 120份样本中,3种试剂阳性率分别为42.50%(51/120)、39.17%(47/120)和41.67%(50/120),阳性检出率无统计学差异(χ²=0.298,P>0.05),对4种样本检测阳性检出率,差异无统计学意义(P>0.05)。3种试剂检测结果一致的有89份。检测结果一致性从高到低为:A试剂和C试剂>B试剂和C试剂>A试剂和B试剂。结论 由于不同厂家试剂灵敏度和准确性不同导致检测结果和检测一致性存在差异,提示试剂厂家需进一步优化性能,提高检测灵敏度和准确性,同时各检测机构应采取多种质控方法以确保新冠病毒核酸检测准确性,以便更好地为疫情防控、病人救治提供科学依据。     

Two cases of HIV-1 acute infection with antibody negative by immunochromatography method

We found two cases of HIV-1 acute infection, confirmed by nucleic amplification test (NAT) and/or RT-PCR, with HIV-1 antibody negative by immunochromatography (IC) method but weakly positive by particle agglutination (PA) test. These cases suggested that IC method was less sensitive than PA test in the detection of acute infections. It is necessary to execute the post counseling that considers the possibility of the acute infection in public health centers and testing places where IC method is used for the screening test. It is also important to recommend taking the following re-examination after a certain period to a person who seems to have had a chance of infection in a short time before testing.     

血吸虫病核酸诊断技术的研究进展

血吸虫病是一种危害严重的重要人兽共患寄生虫病.由于多年积极有效的防治,血吸虫的感染率和感染度明显下降,传统的病原学和免疫学诊断方法已无法满足疫区查病的需要.近年来,国内外研究者们发展了一些针对血吸虫来源DNA的核酸诊断技术,显示了很好的应用前景.该文就核酸检测在血吸虫病诊断中的最新进展进行综述,主要包括靶DNA片段的选...     

Antigen detection, virus culture, polymerase chain reaction, and in vitro antibody production in the diagnosis of vertically transmitted HIV-1 infection

Polymerase chain reaction (PCR), virus culture (V), antigen detection (Ag), and in vitro antibody production (IVAP) assays may be useful for the early detection of vertically transmitted HIV-1 infection in infants under 18 months of age, when a diagnosis cannot be based on seropositivity because of maternal antibody persistence. To assess the reliability of these procedures and to correlate diagnostic results with infection status, 101 children born to HIV-1-seropositive mothers were evaluated by all these techniques within the first 6 months of life. The children were then followed up to the age of at least 18 months, when diagnosis was made on the basis of AIDS or AIDS-related complex (ARC) onset or persistence of HIV-1 seropositivity. Out of 27 children classified as infected according to the above criteria, 25 (92.5%) were repeatedly positive in IVAP test, 22 (81.5%) in the first PCR analysis, and only 19 (70.3%) in the initial V assay. On further testing, a total of 24 children (88.9%) were found positive in PCR assay, and 23 (85.2%) in V test. All these assays were found to be more sensitive than antigen detection for HIV-1 infection diagnosis, but the antigenaemia was shown to be a useful prognostic marker of disease onset. We also found that both Ag and IVAP assays could give false-positive results in the first 2 months of life, which severely limits their diagnostic value during this period of time. False-positive results in PCR assay could occur at any time of the tested period and were unrelated to the child's age.(ABSTRACT TRUNCATED AT 250 WORDS)     

新型冠状病毒抗体检测在疑似病例排查中的应用研究

目的 比较不同原理抗体检测试剂对新冠疑似病例诊断的准确性,对新冠疑似病例提出高效的排查方案,进一步为建立高风险人群的临床筛查路径提供科学依据。方法对76例由于IgM抗体初检阳性被转入定点医院隔离观察的新冠疑似病例,进行鼻咽拭子核酸检测,血清微量病毒中和抗体和3种市售不同原理抗体检测试剂检测,以及肺部CT检查,结合临床症状等信息,综合判断新冠确诊或排除情况,并将3种血清学抗体检测试剂与临床判断结果进行对比分析,比较3种检测方法结果的差异。结果通过综合判断,76例疑似病例中有3例为确诊病例,73例为排除病例。与临床确诊/排除结果比较,3种血清学抗体检测方法中,化学发光总抗体对3例病例入院不同时间点共9份样本均能检出,对73例排除病例入院第1天采集样本的检测特异性为97.26%,高于化学发光法IgM/IgG和胶体金法IgM/IgG单独检测(入院第1 d样本检测特异性均为73.97%)。结论化学发光法检测新冠总抗体具有高灵敏度和高特异性,可以用于疑似病例抗体检测的初筛。针对重点人群筛查,应首先进行2019-nCoV核酸检测,若核酸检测阴性,则采用检测血清总抗体为主要指标的筛查路径。     

Antibodies to the nef protein and to nef peptides in HIV-1-infected seronegative individuals

The silent period that follows infection by the human immunodeficiency virus (HIV-1) and precedes seroconversion remains a problem for the screening of blood supply, and knowledge about the mechanism involved in the maintenance of latency is only fragmentary. Using purified nef recombinant protein and six synthetic nef peptides, antibodies to the product of an HIV-1 regulatory gene, the negative regulatory factor (nef) involved in maintenance of proviral latency, were detected by Western blot and radioimmunoassay techniques in HIV-1-seronegative, viral antigen-negative, and virus culture-negative individuals at risk for HIV infection. This antibody response to nef was correlated in eight individuals with the detection of HIV-1 proviral DNA by oligonucleotide hybridization, following enzymatic amplification of HIV DNA in peripheral blood mononuclear cells. Such latent HIV infections have now been followed for up to 6 or 10 months in five individuals. In addition, retrospective and prospective analysis of HIV-1-seropositive individuals have shown (1) antibodies to nef preceding seroconversion, and (2) the persistence of antibodies to nef and of HIV-1 proviral DNA in a case of spontaneous complete HIV-1 seronegativation. Since DNA amplification cannot be currently considered for routine use, screening for anti-nef antibodies followed by confirmation by DNA amplification could represent a basis for new diagnostic strategies. Beyond their diagnostic implications, these findings, suggesting that regulatory genes of the HIV-1 provirus can be expressed prior to the initiation of virion synthesis, may also be applicable in the design of alternative vaccines against the acquired immunodeficiency syndrome.     

应用全自动核酸检测系统MPLC-COBAS筛查献血员HIV-1 RNA

目的建立一套全自动核酸检测系统筛检献血员艾滋病病毒1型(HIV-1)核糖核酸(RNA)。方法应用MagNAPureLC(MPLC)系统提取核酸,COBASAmplicor系统进行基因扩增与检测,再应用国际标准品和临床标本验证MPLC-COBAS系统。结果该系统的检测灵敏度(95%CI)HIV1RNA病毒为45～67IU/ml,5124份标本混样检测结果为阴性,81份HIV抗体(抗HIV)酶联免疫吸附试验(ELISA)假阳性混样检测结果及6份追踪样本核酸检测结果均为阴性。结论该系统具有全自动检测、灵敏度高、特异性好的特点,可进一步扩大临床标本检测量,探索出切实有效的核酸检测系统进行献血员筛查HIV1RNA。     

Point‐of‐care diagnosis of tuberculosis: Past,present and future

Diagnosis represents only one aspect of tuberculosis (TB) control but is perhaps one of the most challenging. The drawbacks of current tools highlight several unmet needs in TB diagnosis, that is, necessity for accuracy, rapidity of diagnosis, affordability, simplicity and the ability to generate same‐day results at point‐of‐care (POC). When a return visit is required to access test results, time to treatment is prolonged, and default rates are significant. However, a good diagnostic tool is also critically dependent on obtaining an adequate biological sample. Here, we review the accuracy and potential impact of established and newer potential POC diagnostic tests for TB, including smear microscopy, the Xpert MTB/RIF assay (Cepheid) and the Determine TB lipoarabinomannan antigen test (Alere). Novel experimental approaches and detection technologies for POC diagnosis of active TB, including nucleic acid amplification tests, detection of volatile organic compounds or metabolites, mass spectroscopy, microfluidics, surface‐enhanced Raman spectroscopy, electrochemical approaches, and aptamers among others, are discussed. We also discuss future applications, including the potential POC diagnosis of drug‐resistant TB and presumed latent TB infection. Challenges to the development and roll‐out of POC tests for TB are also reviewed.