双抗体酶联免疫法测定血小板抗体

双抗体酶联免疫法测定血小板抗体太原市中心医院（０３０００９）辛秋绒，李翠香，贾玉萍，史玉亮我们从１９９１年开始用酶联免疫法对免疫性血小板减少疾患和非免疫性血小板减少疾患的血小板抗体（ＰＡＩｇＧ）进行了检测，并与健康人做了对照。现将检测结果报告如下。对...     

化学发光法和酶联免疫法在丙型肝炎病毒抗体检测中的应用价值

目的探讨化学发光法和酶联免疫法(ELISA)在丙型肝炎病毒(HCV)抗体检测中的应用价值。方法研究时间为2016年1月至2017年9月,选择在我院进行丙型肝炎检测的患者26456例作为研究对象,取所有患者的血清样本,采用化学发光法和酶联免疫法对抗HCV抗体进行检测,判定阳性情况。结果化学发光法检测抗HCV抗体阳性345例,阴性26111例,检出率为1.30%;ELISA法检测抗HCV抗体阳性350例,阴性26106例,检出率为1.32%,对比无显著差异(P>0.05)。两种方法共同检出阳性样本344例,ELISA法检测阳性而化学发光法检测阴性为1例,化学发光法检测阳性而ELISA法检测阴性的有6例。结论相对于ELISA法,化学发光法在丙型肝炎病毒抗体检测中的应用具有更好的敏感性,有很好的应用价值。     

国家食品药品监督管理局对谷氨酸脱羟酶抗体酶联免疫法检测试剂等产品作出分类界定

为适应医疗器械监督管理需要，日前，国家食品药品监督管理局发出通知，对谷氨酸脱羟酶抗体酶联免疫法检测试剂等两种诊断试剂的作出分类界定。     

酶免疫法检测丙型肝炎病毒血清型

采用丙型肝炎病毒核心区及NS4区合成肽键建立酶联免疫检测方法,对血清中丙型肝炎病毒抗体进行分型。测定抗-HCV阳性者再测其血清型。对HCV RNA阳性的标本用特异性引物的分型PCR方法测定基因型。结果表明,70例供血者中HCV RNA阳性为30％,抗-HCV阳性率68．6％,其中血清分型阳性率为72．9％。血清Ⅰ型与基因Ⅱ（1b）型相对应;血清2型与基因Ⅲ（2a）型相对应。     

酶联免疫法检测血清中抗PPD抗体对结核病诊断的价值

我所近年来采用ELISA法检测94例结核与非结核患者血清中抗PPD抗体，报道如下。材料和方法一、对象所有病例均为1993年6月～1994年3月到本所就诊的门诊病人。1．活动性肺结核组：49例，男肠例，女13例；年龄423±15．6岁，其中菌阳16例。2．非活动性肺结核组：31例，男22例，女9     

酶联免疫吸附试验加样量对丙型肝炎病毒抗体检测的影响

酶联免疫吸附试验(ELISA)检测抗丙型肝炎病毒(HCV)抗体目前已是血站、医院的常规检测项目,虽然该方法操作简单、灵敏度高,但在实际操作中,ELISA检测抗-HCV抗体结果假阳性、假阴性问题还是比较常见,这给临     

汉坦病毒(流行性出血热病毒)抗体IgM酶联免疫试剂应用评价

本文应用汉坦现毒抗体IgM酶联免疫试剂对192例EHF病人和疑诊患者39例进行EHF抗体IgM检测,检测结果提示153例EHF病人血清有148例,阳性检出率为96.7%（148／153）;39例疑诊病人血清IgM均为阴性,二者有明显差异,P＜0.01。其中有两例疑诊病人在一周内先后三次进行EHF－IgM检测都为阴性,说明该法灵敏度高,特异性强,重复性好的优点,适合于基层医疗单位汉坦病毒感染流行性病学调查。     

化学发光法和酶联免疫吸附法检测丙型肝炎病毒抗体的观察与对比

目的分析和对比应用化学发光法(CLIA)和酶联免疫吸附法(ELISA)对检测丙型肝炎病毒抗体(抗-HCV)准检率和临床意义。方法收集临床血清标本906例,分别应用化学发光法(CLIA)和酶联免疫吸附法(ELISA)监测,并收集监测后阳性标本和可疑阳性标本。最后用第三代重组免疫印迹法(RIBA-3)进行确定再监测。结果两种方法共筛选出血清标本阳性者43例,可疑阳性标本4例。其中CLIA筛选监测出血清阳性者21例,确证为20例,1例可疑阳性。后经RIBA重新确认监测为阳性。敏感性100%。ELISA筛选监测出血清阳性为22例,阴性者3例,确证后阳性为19例,后经RIBA重新确认监测后1例为阴性,2例为阳性。敏感性为95%。结论 CLIA监测法较ELISA监测法灵敏、准确,使用CLIA提高了临床监测率,减少了漏诊现象。对于CLIA和ELISA监测法均显示可疑时,一定要应用RIBA再确诊定性。     

乙肝两对半快速检测板与酶联免疫法的探讨

目的　探讨乙肝两对半检测板在急诊检验中的应用价值。方法　用金标检测试剂板测定血清 (血浆 )中乙肝五项标志物 ,结果与 EL ISA法相比较。结果　 2 74例标本中 ,用 EL ISA法与金标法有 2 73例一致 ,EL ISA法有2 0 4例阳性 ,70例阴性 ,金标检测试剂板有 1例 HBs Ag阳性未能检出 ,其余各例两法结果完全符合。结论　金标法与酶联法测乙肝两对半的符合率高 ,结果基本一致 ,且金标法快速简单 ,更适合于急诊检验     

酶联免疫吸附试验双抗体夹心法检测丙型肝炎病毒抗原的临床符合性研究

目的探讨丙型肝炎病毒抗原酶联免疫吸附试验(ELISA)检测试剂盒的临床符合性。方法采用科华公司HCV抗体ELISA检测试剂盒和HCV RNA基因扩增试验作为对照。结果对照两种方法,其敏感性均为100N,特异性分别为92％和90％,类风湿(RF)阳性血清无干扰,阳性和阴性血清的批内(n-10)变异系数分别为8．15％和11．08％。结论丙型肝炎抗原ELISA检测方法敏感性高,特异性好,操作简便易行,不需特殊的仪器设备、费用低廉,具有推广前景。     

Small-molecule inhibitors of hepatitis C virus (HCV) non-structural protein 5A (NS5A): a patent review (2010-2015)

Introduction: Non-structural 5A (NS5A) protein has achieved a considerable attention as an attractive target for the treatment of hepatitis C (HCV). A number of novel NS5A inhibitors have been reported to date. Several drugs having favorable ADME properties and mild side effects were launched into the pharmaceutical market. For instance, daclatasvir was launched in 2014, elbasvir is currently undergoing registration, ledipasvir was launched in 2014 as a fixed-dose combination with sofosbuvir (NS5B inhibitor).

Areas covered: Thomson integrity database and SciFinder database were used as a valuable source to collect the patents on small-molecule NS5A inhibitors. All the structures were ranked by the date of priority. Patent holder and antiviral activity for each scaffold claimed were summarized and presented in a convenient manner. A particular focus was placed on the best-in-class bis-pyrrolidine-containing NS5A inhibitors.

Expert opinion: Several first generation NS5A inhibitors have recently progressed into advanced clinical trials and showed superior efficacy in reducing viral load in infected subjects. Therapy schemes of using these agents in combination with other established antiviral drugs with complementary mechanisms of action can address the emergence of resistance and poor therapeutic outcome frequently attributed to antiviral drugs.     

Studies on the development of enzyme linked immunosorbent assay (ELISA) for hepatitis B surface antigen(HBsAg) by monoclonal antibodies of different affinity constants

Mouse monoclonal antibodies to Hepatitis B surface antigen(HBsAg) were prepared and their functional capabilities tested by the method of solid phase enzyme linked immuno sorbent assay(ELISA). HBsAg binding studies indicated that one monoclonal antibody 6E-1-1 bound more HBsAg at a faster rate than the other monoclonal antibodies. Also, for the binding inhibition studies with the selected monoclonal antibody 6E-1-1, one monoclonal antibody 8D-3-6 didn’t exhibit binding inhibition for HBsAg. Then, a simultaneous ELISA method was developed for the immunodiagnosis of HBsAg. Different combinations of two monoclonal antibodies as solid phase and horseradish peroxidase(HRPO) labeled phase were studied. The combination of monoclonal antibody of higher affinity constant (6E-1-1) immobilized in a solid phase and monoclonal antibody of lower affinity constant (8D-3-6) as a HRPO labeled phase was more sensitive when two monoclonal antibodies of different affinity constants for HBsAg were prepared.     

重组蝎毒抗神经兴奋肽的ELISA分析方法建立及稳定性的初步研究

目的建立重组蝎毒抗神经兴奋肽(ANEP)的ELISA分析方法并对ANEP的稳定性进行初步研究。方法以分别建立的间接ELISA和间接竞争ELISA分析方法进行ANEP含量测定,并通过方法学的优化,最终以建立的间接法ELISA方法测定了温度、pH值、反复冻融、超声对ANEP稳定性的影响。结果间接ELISA相对于间接竞争ELISA有着更好的线性关系和灵敏度,线性范围0.025~1.60μg/mL。检测限为10 ng/mL。稳定性实验表明ANEP在37,60℃的条件下放置24 h含量基本不变,pH 5~11的缓冲液中保持稳定,反复冻融、超声4 min后含量降低。结论所建立的间接ELISA方法能很好的用于ANEP的测定。ANEP的热稳定性较好,强酸性下不稳定,对反复冻融与超声等稳定性较差。     

The Shift in Emphasis From Risk-Based to Age-Based Hepatitis C Virus (HCV) Testing in the US Tends to Remove Injection Drug Use From Discourse on HCV

Background: Hepatitis C virus (HCV) infection is hyperendemic among people who inject drugs; nonsterile drug injection is the principle risk for HCV acquisition. Due to gaps in the HCV care continuum, there have been recommendations in the United States emphasizing age—rather than risk-based testing strategies. The central research focus of this project is to explore the meanings and implications of the shift in emphasis from risk-based to age-based HCV testing with regard to people who use drugs. Methods: Content analysis and critical discourse analysis, informed by eco-social theory, were used to examine relevant documents. Results: Fifteen documents were assessed for eligibility; 6 documents comprised the final set reviewed. In content analysis, age-based testing was both mentioned more frequently and was supported more strongly than risk-based testing. Risk-based testing was frequently mentioned in terms minimizing its use and drug use was often mentioned only euphemistically. The reframed emphasis largely removed discussion of injection drug use from discussion of HCV risks. Conclusion: Shifting the emphasis of HCV testing from testing based on specific routes of transmission and risk to testing based on age removes injection drug use from HCV discourse. This has the potential to either facilitate HCV care for drug users or to further stigmatize and marginalize drug use and people who use drugs. The potential implications of this shift in testing emphasis for public health merit further investigation.     

Lipophilic derivatization of synthetic peptides belonging to NS3 and E2 proteins of GB virus‐C (hepatitis G virus) and its effect on the interaction with model lipid membranes

Abstract: The synthesis by solid‐phase methodologies of peptides belonging to structural and non‐structural proteins of GB virus C as well as its N‐α‐acylation with myristate and palmitate fatty acids is described. To explore the peptide–lipid interactions we have used liposomes composed of dipalmitoylphosphatidylcholine as model membranes and complementary spectroscopic and calorimetric techniques. Our results show that structural and more clearly the structural lipophilic peptide sequences incorporated into lipid bilayers perturb the packing of lipids and affect their thermotropic properties, more than the non‐structural selected sequence. However, the binding of the synthetic sequences to lipid membranes occurred without any restructuration of the peptides.     

The effect of cytokine profiles on the viral response to re-treatment in antiviral-experienced patients with chronic hepatitis C virus infection

Background

There have been few studies on the potential immunological factors associated with viral controls in antiviral-experienced patients on a second round of combination therapy. In this study, we evaluated the level of systemic cytokines and potential impact on combination therapy in both antiviral-naïve and -experienced patients chronically infected with hepatitis C virus.

Methods

Longitudinal analysis of 27 cytokines and chemokines was performed using the multiplex Biorad 27 plex assay in 37 antiviral-naïve and 24 experienced chronically HCV-1b-infected patients during combination therapy with peginterferon-alfa and ribavirin. A group of healthy donors was included as the control (n = 11).

Results

Fifty percent of antiviral-experienced chronically HCV-patients could achieve a delayed and slow virologic response after 48 weeks combination therapy, comparing with an early and fast virologic response in antiviral-naïve patients. A distinction of immune mediators profiling before and during antiviral therapy between antiviral-naïve and -experienced patients was identified, IL-4, IFN-γ and CCL-3 (MIP-1a) were significantly higher in naïve patients than those in experienced patients (P = 0.005, 0.047 and 0.017, respectively) while G-CSF in naïve was lower than in experienced patients (P < 0.05). Notably, higher Th1 type cytokine IFN-γ and lower Th2 type cytokine IL-4 at baseline and week 4 were associated with HCV clearance in naïve patients, and a similar trend appeared at week 12 in experienced patients.

Conclusions

We found a successful second round therapy in antiviral-experienced patients appears to be associated with the host immune response. Dominant Th1-polar cytokines, especially IFN-γ, is a potential predictor of viral responsiveness.