The detection of infectious mononucleosis antibodies by radioimmunoassay and ELISA

An infectious mononucleosis receptor (IMR) preparation has been isolated from sheep erythrocyte stroma. This material was used to detect the presence of human infectious mononucleosis (IM) antibodies with rabbit anti-human immunoglobulin antisera by radioimmunoassay and ELISA. These procedures use 10-20 ng of IMR and 5-10 microliter of human serum. A single treatment of the IMR with neuraminidase reduces its ability to bind to the IM antibodies by 80%. In addition absorption of the IM sera with sheep red cells completely removes IM specific antibodies.     

An improved method for monitoring efficacy of anti-retroviral therapy in HIV-infected individuals: a highly sensitive HIV p24 antigen assay.

Circulating human immunodeficiency virus (HIV) p24 antigen levels were measured by a highly sensitive HIV p24 antigen-capture enzyme-linked immunosorbent assay (ELISA) in patients with acquired immunodeficiency syndrome (AIDS) and AIDS-related complex (ARC) otherwise negative for HIV p24 antigen measured by a commercial antigen-capture ELISA. The assays were performed at baseline and at several intervals during treatment with either zidovudine (ZDV) or dideoxyinosine (ddl). To further enhance the rate of antigen detection, serum was pretreated with hydrochloric acid to denature antibody in immune complexes. Utilizing this assay system, we monitored these patients for drug efficacy. HIV p24 antigen levels obtained by using this sensitive assay decreased in 3 of 8 patients receiving ZDV during 8 weeks of ZDV treatment. Similarly, ddl administration was associated with a decrease of HIV p24 antigen levels in 3 of 5 patients. Thus, the use of the highly sensitive HIV p24 antigen assay permitted the monitoring of surrogate HIV p24 antigen as a measure of efficacy of anti-retroviral therapy in all of these patients who were otherwise HIV p24 antigen-negative at the onset of anti-retroviral therapy.     

ELISA及免疫印迹法检测抗肌球蛋白抗体的比较

目的 比较ELISA和免疫印迹法检测抗肌球蛋白抗体的优缺点和临床应用价值。方法 采用猪心肌组织中提取的心肌肌球蛋白作抗原,以ELISA和免疫印迹法检测48例扩张型心肌病（DCM）和40例冠心病患者血清中的抗肌球蛋白抗体;同时与人工合成的肌球蛋白重链抗原对比。结果 ELISA和免疫印迹法对DCM组抗肌球蛋白抗体检测的阳性率相近（50％对43．8％）,但后者能区分针对抗原亚单位的抗体,前者则不能。猪心肌肌球蛋白与人工合成多肽的ELISA检测结果具有高度的相关性（P＜0．01）。结论 以人工合成肌球蛋白重链作抗原,采用ELISA检测抗肌球蛋白抗体既有助于DCM与冠心病的鉴别诊断,又简便易行。     

Follow-up study of individuals with high tetraplegia (C1-C4) 14 to 24 years postinjury

OBJECTIVE: To determine the life circumstances and psychosocial status of individuals with respirator-assisted and respirator-independent high tetraplegia an average of 19 years after spinal cord injury. DESIGN: Survey data were analyzed separately for ventilator-assisted and ventilator-independent groups. SETTING: Three spinal cord injury rehabilitation centers in California, Colorado, and Texas. SUBJECTS: Eighty-two individuals with CI-C4 tetraplegia between 14 and 24 years postinjury who had received acute inpatient rehabilitation. MAIN OUTCOME MEASURES: Demographics, health care utilization patterns, activities of daily living (Katz Level of Free Time Activities Scale), self esteem (Rosenberg Self Esteem Scale), quality of life, and employment. RESULTS: Self esteem and quality of life were, reported as high. Most subjects had some form of health care insurance. More than 90% lived in private homes. Approximately one third of cases had at least a college degree, yet only one quarter reported being employed. One fifth of individuals were married. Almost half of ventilator-independent cases and one quarter of ventilator-assisted cases lived in households with income of less than $20,000 per year. Mean hospital days in the past year were 11 for the ventilator-independent group and 6 for the ventilator-assisted group. The latter group required more nursing level care, significantly more hours of care, and more paid attendants over the year. Ninety-five percent of individuals reported being "glad to be alive." CONCLUSIONS: Assistance in the areas of socialization, financial status, personal assistance services, transportation, and entry into competitive employment were defined as needed. Quality of life was higher than expected, considering the substantial physical limitations of the group. The sample was almost unanimously glad to be alive, including all ventilator-assisted individuals.     

Intrathecally produced antibodies to Borrelia burgdorferi measured by IgG-capture ELISA

An IgG-capture enzyme-linked immunosorbent assay (ELISA) which directly measures the intrathecal production of Borrelia burgdorferi-specific IgG in neuroborreliosis is described. The method is based on the simultaneous measurement of IgG-specific antibodies in paired cerebrospinal fluid (CSF) and serum samples. Sixty-nine paired CSF/serum samples from 64 patients with neuroborreliosis or other neurological disorders and six samples from acrodermatitis chronica atrophicans (ACA) patients were analysed by IgG-capture ELISA. Of 12 samples from patients with early neuroborreliosis, intrathecal production of B. burgdorferi-specific IgG could be demonstrated in 11 samples. Of 56 patients with other neurological disorders of known or unknown etiology, intrathecal production of B. burgdorferi-specific IgG was demonstrated in one patient who had a history compatible with a previous neuroborreliosis. No intrathecal production was found in the samples from ACA patients in spite of high Borrelia titres in their CSF as measured by indirect ELISA. In conclusion, the B. burgdorferi IgG-capture ELISA is a sensitive and reliable method for the detection of intrathecal production of IgG antibodies to B. burgdorferi. It is advantageous in comparison to the indirect ELISA as it does not require correction for leakage across the blood/CSF barrier.     

Late presentation of HIV-infected individuals

Late presentation remains a major concern despite the dramatically improved prognosis realized by ART. We define a first presentation for HIV care during the course of HIV infection as 'late' if an AIDS-defining opportunistic disease is apparent, or if CD4+ T-cells are <200/microl. In the Western world, approximately 10 and 30% of HIV-infected individuals still present with CD4+ T-cells <50 and <200/microl, respectively; estimates are substantially higher for developing countries. Diagnosis and treatment of opportunistic diseases and intense supportive in-hospital care take precedence over ART. Benefits of starting ART without delay, that is, when opportunistic diseases are still active, include faster resolution of opportunistic diseases and a decreased risk of recurrence. The downside of starting ART without delay could include toxicity, drug interactions and immune reconstitution inflammatory syndrome (IRIS). Among asymptomatic or oligosymptomatic individuals presenting late, where ART and primary prophylaxis are initiated, approximately 10-20% will become symptomatic from drug toxicity or undiagnosed opportunistic complications, including IRIS, which require appropriate therapies. In this review we describe late presentation to HIV care, the scale of the problem, the evaluation of a late-presenting patient and challenges associated with initiation of potent antiretroviral therapy (ART) in the setting of acute opportunistic infections and other comorbidities.     

ELISA检测血清抗磷脂酶A_2受体抗体在膜性肾病中的应用探讨

目的探讨酶联免疫吸附试验(ELISA)检测血清抗磷脂酶A2受体抗体(抗PLA2R抗体)滴度在膜性肾病中的应用价值为疾病诊断及病情判断提供更特异的血清学指标。方法选取该院肾内科就诊行肾组织活检并确诊的膜性肾病患者34例,该院住院自身免疫性疾病及肾病综合征、肾功能不全患者32例,健康体检者12例,收集血清检测抗PLA2R抗体并结合血清总蛋白(TP)、清蛋白(ALB)、IgG、IgA、IgM、C3、C4指标,分析其对膜性肾病的诊断性能。结果抗PLA2R抗体滴度在膜性肾病组、疾病对照组、健康对照组中位数依次为22.1、2.0、2.0RU/mL,膜性肾病组与其他两组间比较差异有统计学意义(P0.05)。抗PLA2R抗体在膜性肾病组阳性17例(阳性率50%),阴性17例,疾病对照组及健康对照组均为阴性,其特异度与阳性预测值为100%。膜性肾病组TP、ALB、IgG与疾病对照组、健康对照组比较差异有统计学意义(P0.05);抗PLA2R抗体与血清TP、ALB、IgG、IgA、IgM、C3、C4的相关系数依次为-0.382、-0.344、-0.502、-0.295、0.062、0.005、0.241,与TP、ALB、IgG、IgA呈负相关(P0.01),与C4呈正相关(P0.05),与IgM、C3无相关性(P0.05)。结论抗PLA2R抗体在膜性肾病诊断中具有较高的特异性,可作为无法行肾活检患者的必要和特异性血清学检测指标,定量检测有助于病情判断。     

Stavudine, lamivudine plus novel protease inhibitor therapy in antiretroviral-naive HIV-infected individuals treated for 24 weeks

Preliminary results are presented from a dose-comparison trial of the regimen stavudine/lamivudine plus the novel protease inhibitor, ABT-378/ritonavir, given to 101 antiretroviral-naive, human immunodeficiency virus (HIV)-infected subjects for > or = 24 weeks. The HIV-1 RNA had decreased to <400 copies/ml in 94% of patients and CD4 cell count had increased by approximately 160 cells/mm3 at 24 weeks. The regimen was well tolerated and merits further study.     

Determination of the level of antibodies to CD4 in serum of patients with malignant lymphomas and of HIV-infected individuals

Hardphase ELISA was used to compare the blood sera of patients with malignant skin lymphomas and HIV infection and of healthy volunteers for autoimmune antibodies to CD4. Antibodies to CD4 were not detected in the volunteers (30 sera) and in the patients with malignant skin lymphomas (52 patients with different disease stages). The antibodies were found in 4 of 11 HIV cases.     

Development of a comparative dot ELISA for the detection of antibodies in blastomycosis

We present a preliminary study utilizing an enzyme immunoassay for the detection of antibodies in blastomycosis using nitrocellulose membrane as the solid support. The comparative dot enzyme-linked immunosorbent assay (CDE) utilizing Blastomyces dermatitidis, Histoplasma capsulatum, and Coccidioides immitis antigens allows rapid screening of specimens and determination of a serum profile with respect to anti-Blastomyces antibodies. Sensitivity (76% or greater) and specificity (approximately 12% crossreactivity) determinations indicated that this visual assay may be an alternative antibody detection method for blastomycosis or other systemic fungal diseases.     

Development and application of a GuHCl-modified ELISA to measure the avidity of anti-HPV L1 VLP antibodies in vaccinated individuals

Antibody responses against infectious agents are an important component in the prevention of disease. The avidity of antibodies for their antigens relates to their functional efficiency, and is a fundamental aspect in the investigation of humoral responses. Modified ELISAs are used to estimate avidity through the use of chaotropic agents and the measurement of the degree to which they disrupt the interaction between antibody and antigen. The theory behind the assay is the higher the avidity of an interaction the less susceptible it is to the effects of the chaotropic agent. The goal of this study was to generate a modified ELISA where a complex, multimeric coating-antigen, human papillomavirus (HPV) virus-like particles (VLP), was used to measure the avidity of anti-HPV antibodies generated following vaccination with HPV VLPs. A series of chaotropic agents were evaluated in the assay for their effectiveness in measuring avidity. Guanidine hydrochloride (GuHCl) was selected as a chaotropic reagent with the ability to disrupt antibody and antigen interactions, while not affecting the integrity of the plate-bound VLP. Two methods of determining the avidity index were assessed and shown to be comparable. This assay was then successfully applied to measure the avidity of anti-HPV VLP serum antibodies in samples from an HPV L1 VLP vaccine clinical trial. Overall, the assay was highly reproducible and captured a wide range of antibody avidities. Therefore, a GuHCl-modified ELISA is an acceptable method that can be used to determine HPV-specific antibody avidity indices within a clinical trial setting.     

Newly developed Norwalk-like viruses detection ELISA by combination with monoclonal antibodies and hyperimmune rabbit sera

Norwalk-like viruses(NLVs) antigen detection ELISA was newly developed. It is constructed with monoclonal antibodies(MAbs) as a capture antibodies and hyperimmune rabbit sera as a detector antibodies. The two MAbs reacted broadly and specifically with genogroup I and genogroup II NLVs, respectively. Total detection time is less than 3 hours and at least 90 stool samples could be tested at one time. The concordance rate with ELISA and RT-PCR was 69% and it could be raised to 71% if capsid primer sets were used. Epidemiological study revealed that among 664 normal persons one sample(0.2%) was confirmed positive by both ELISA and RT-PCR, suggested that NLV carrier may exist in the normal person. The ELISA kit has advantages such as rapid, simple and multi samples detection, and useful for the first screening test. Also it is big tool for epidemiological surveillance of NLVs invasions.     

Characterization of antibodies produced by S-s- individuals

BACKGROUND: Historically, classification of U- and U variant (U+var) individuals has been made by hemagglutination and adsorption and elution studies performed with polyclonal U antisera. Molecular studies and serologic tests with a potent monoclonal anti-He have shown that U+var red cells, some of which are He+, possess an altered form of glycophorin B. STUDY DESIGN AND METHODS: Seventeen sera, previously determined to contain anti-U, were tested with a panel of red cells of common and rare MNS types. RESULTS: Five sera contained anti-U only, and 12 sera contained broadly reactive antibodies with apparent, but inseparable, anti-U,He or anti-U,N,He specificities. CONCLUSION: The majority of antibodies produced by S-s-U- individuals are anti-U plus anti-glycophorin B and are analogous to the broadly reactive antibodies produced by En(a-) individuals whose red cells lack glycophorin A or have altered glycophorin A. To avoid further immunization of patients with anti-U, sera used for classification of S-s-U- donors should be selected to detect S-s- red cells that possess altered forms of glycophorin B.     

The respiratory microbiome of HIV-infected individuals

Introduction: The respiratory tract is constantly exposed to various environmental and endogenous microbes; however, unlike other similar mucosal surfaces, there has been limited investigation of the microbiome of the respiratory tract.

Areas covered: In this review, we summarize the current state of knowledge of the bacterial, fungal, and viral respiratory microbiomes during HIV infection and how the microbiome might relate to HIV-associated lung disease.

Expert commentary: HIV infection is associated with alterations in the respiratory microbiome. The clinical implications of lung microbial dysbiosis are however currently unknown. Mechanistic studies are needed to establish causality between shifts in the respiratory microbiome and pulmonary complications in HIV-infected individuals.     

双抗原夹心ELISA检测抗HCV总抗体

目的建立检测抗HCV抗体的双抗原夹心ELISA法,评价其可行性。方法将与His或GST融合表达的HCV基因工程抗原,分别用作ELISA的包被和酶标记抗原,建立用于抗HCV总抗体检测的双抗原夹心ELISA。用此方法检测1 968份临床血清标本,并以间接ELISA(北京万泰试剂)与之对照;此外,用套式RT-PCR检测部分血清的HCV RNA。结果有1 761份血清2种ELISA检测均为阴性,有190份血清均为阳性,两种方法符合率为99.1%;有17份血清的检测结果不相符,间接法阳性而本法阴性的14份,其中HCV RT-PCR阳性1份;本法阳性而间接ELISA阴性的3份,其中RT-PCR阳性2份。双抗原夹心ELISA与间接ELISA的敏感性分别为99.48%、98.96%,特异性分别为99.94%、99.27%。结论新研制的检测抗HCV总抗体的双抗原夹心ELISA具有较高的敏感性和特异性,值得作进一步的深入研究。