斑点酶免疫试验检测流行性出血热抗体的研究

本文应用斑点酶免疫试验(DEIA),检测流行性出血热(EHF)病人血清42份,其结果与间接免疫荧光(IFA)进行了比较,符合率100%,其DEIA 几何平均滴度为1:880,IFA 为1:580,与类风湿因子阳性血清[RE( )]无交叉反应,证明本方法具有较高的敏感性和特异性。本文还对不同方法处理组织内源酶的效果和不同阻断剂的阻断效果进行了比较。试验结果表明本法不但敏感、特异,而且简单、快速、安全、经济、易于推广。     

乙型肝炎检测系统的敏感性与性能的评价

检测HBsAg的第三代方法目前包括放射免疫试验(RIA)、反相被动血凝试验(RPHA)和酶标记免疫试验(EIA)。此外,尚有反相被动胶乳凝集试验(RPLA)待批准使用。这些方法的敏感性较第二代方法,如对流免疫电泳高100倍,较第一代方法——琼脂扩散高1000倍。作者根据2,000余份临床血标本检验所取得的经验,对第三代各系统的优缺点进行评价。试验方法简述如下:(1)RPLA:待检标本可预先用吸收试剂处理,以减少类风湿因子或其它非特异性因子引起的假阳性,但试     

酶联免疫法筛查人类免疫缺陷病毒抗体在艾滋病诊断中的应用

目的探讨酶联免疫法筛查人类免疫缺陷病毒(HIV)抗体在艾滋病诊断中的应用效果。方法选择2019年1—12月于医院接受HIV抗体筛查的1000例艾滋病疑似患者作为研究对象,所有患者均行酶联免疫法与金标斑点法检测,以初筛及印证试验联合诊断结果为金标准,比较酶联免疫法与金标斑点法的诊断准确度、灵敏度、特异度。结果酶联免疫法的诊断准确度为99.90%,高于金标斑点法的98.90%,差异有统计学意义(P<0.05);酶联免疫法的诊断灵敏度及特异度分别为100.00%、99.90%,高于金标斑点法的75.00%、99.00%,差异有统计学意义(P<0.05)。结论在艾滋病诊断中应用酶联免疫法筛查HIV抗体具有较高的应用价值,可为临床诊断提供有效依据。     

血清MMP-3、TIMP-1在类风湿关节炎中作用的探讨

目的探讨血清基质金属蛋白酶-3（MMP-3）、基质金属蛋白酶组织抑制剂-1（TIMP-1）在类风湿关节炎（RA）中的作用以及与类风湿因子（RF）的相关性。方法选择44名类风湿性关节炎患者，以酶联免疫吸附试验检测血清MMP-3和TIMP-1，魏氏法测定血沉，免疫透射法测定类风湿因子，并进行结果分析。结果类风湿性关节炎患者的血清MMP-3和TIMP-1含量明显高于正常对照组，差异有统计学意义（P〈0．05），且与类风湿因子、血沉成正相关。结论血清MMP-3、TIMP-1可作为反映类风湿性关节炎病情活动的新指标，比传统的检测指标类风湿因子具有更高的灵敏度。     

末梢血C-反应蛋白胶乳快速测定法

目的 建立C-反应蛋白（CRP）胶乳试验的快速测定法。方法 用末梢血代替静脉血，化学灭活代替加热灭活，改良前处理过程。结果 该法与原胶乳法对照平行试验128例，结果符合率为100％，10例抗类风湿因子（RF）阳性标本，用该法稀释后测定CRP均为阴性。结论 该法快速、稳定，不受RF干扰。     

防病强身

类风湿因子阳性就是得了＂类风关＂吗？ 我年过花甲,平时身体健康,腰腿不酸痛,全身关节也不肿胀。可是我在最近体检时发现类风湿因子阳性。请问我是否得了类风湿关节炎呢？广西徐爱华徐爱华读者：类风湿关节炎（简称类风关）是一种常见的免疫性疾病,一般认为与感染,内分泌紊乱以及免疫功能异常有关。     

免疫斑点技术

免疫酶斑点试验有两种方法,第一种方法为免疫酶电转移技术,文献中也称“西方印染技术”(Westein—bloting),这种方法1979年是美国科学院Towbin首先建立的(Proc.Natl.Acad.SciUSA 1979.76.4350)。另外一种方法是免疫酶斑点试验(Dot Inmunoerzynre assay)是1982年Hawkes建立的(Anal Biochem.1982.119.142)。本期刊有免疫酶斑点试验文章,请参阅。     

两种肺炎衣原体检测方法的比较研究

目的对比微量免疫荧光法和酶联免疫法的敏感性、特异性及抗干扰性,以期找到适合临床推广的方法。方法分别采用微量免疫荧光法和酶联免疫法检测肺炎衣原体IgM抗体(CPN-IgM),包括170份疑似肺炎患儿血清、162份儿童保健科健康体检儿童血清及类风湿因子阳性血清、自身免疫性疾病患者血清、肺炎支原体抗体阳性血清各5份,将结果进行对比研究。结果酶联免疫法CPN-IgM抗体诊断试剂盒与微量免疫荧光检测试剂盒相比,诊断敏感性达90.0%,特异性达94.8%,总的诊断效率达94.2%;且与特殊样本均无交叉反应。结论酶联免疫法CPN-IgM抗体诊断试剂适合在临床检测中推广应用。     

斑点金免疫渗滤试验在流动采血中初筛HBsAg的探讨

实验室检测献血者HBsAg一般采用酶联免疫吸附法(ELISA) [1] ,其实验条件要求严格 ,操作程序较为复杂 ,费时较长 ,不适应流动采血。为适应街头无偿献血工作需要 ,笔者引用斑点金免疫渗滤试验对献血者进行HBsAg初筛 ,并与ELISA方法检测HBsAg结果作比较 ,并对HBsAg斑点金免疫渗滤试验检测试剂试验条件与假阴性方面进行了观察 ,现报道如下。1　材料与方法1.1　材料　HBsAg斑点金免疫渗滤试验试剂 ,郑州博赛生物工程有限责任公司出品 ,批号 2 0 0 10 10 2 ,灵敏度 :1ng/ml ;ELISA法试剂 :厦门新创科技有限公司出品 ,批号 2 0 0 10 2 …     

斑点酶联免疫吸附试验在诊断滴虫性阴道炎的临床应用

目的 探讨不受温度影响且较准确诊断滴虫性阴道炎的方法。方法 采用斑点酶联免疫吸附试验。结果 检测１０８例经生理盐水涂片检查滴虫阳性者标本，其抗体阳性率为９２．５％，检测真菌阳性者标本５５例，滴虫抗体阳性率为０。结论 斑点酶联免疫吸附试验是一种灵敏度高、重复性好、特异性强、不受温度影响的诊断滴虫性阴道炎的方法。     

卡马西平血清样品的稳定性考察

目的考察卡马西平血清样品不同保存条件对血清药物浓度的影响。方法取癫痫患者血清样品,在取血当时、室温(28±3)℃保存1d、2d、3d或冷藏(4±1)℃保存1d、3d、7d后,用荧光偏振免疫法(FPIA)测定血清卡马西平浓度并进行比较,考察其稳定性。结果室温保存1d、2d、3d或冰箱冷藏保存1d、3d、7d的血清测定结果均与取血当时测定结果无统计学差异。结论卡马西平血清样品在室温下可稳定保存三天、冷藏条件下可稳定保存七天。     

抗环瓜氨酸肽抗体检测在类风湿关节炎诊断中的应用

目的：探讨抗环瓜氨酸肽（抗-CCP）抗体对类风湿关节炎（RA）的临床应用价值,并比较与类风湿因子（RF）之间的相关性以及在临床诊断上的意义。方法：对75例RA患者血清标本和77例对照者的血清标本用酶联免疫吸附试验（ELISA）进行抗-CCP抗体检测,并测定RF,分析RF与抗-CCP抗体的相关性。结果：75例RA患者中,抗-CCP抗体阳性率为56%（42/75）,非RA对照组的阳性率为5.2%（4/77）,两者有显著差别。抗-CCP抗体对RA诊断的敏感度为56%,特异性为94.8%;阳性和阴性似然比分别为16.08和0.16。抗-CCP抗体和类风湿因子（RF）具有相关性。结论：抗-CCP抗体对RA诊断具有良好的敏感度和特异性,将是协助诊断RA的一个新的实验指标。     

清远市辖监狱服刑人员艾滋病感染情况调查

目的 了解清远市辖监狱服刑人员艾滋病感染情况,为预防、控制和治疗提供参考依据。方法 对清远市辖监狱（包括清远监狱和英德监狱）的服刑人员进行全员调查,询问其危险行为史并抽取血液样本,采用双抗原夹心酶联免疫法进行HIV抗体筛查试验,筛查结果阳性的样本送广东省HIV确认中心实验室,采用蛋白印迹法进行确认,并报出结果。结果 在14862名服刑人员血样中检测出HIV抗体阳性61例,总体阳性率0.41%;其中有42名服刑人员有静脉吸毒史,占HIV抗体阳性人数的68.85%;在4275名清远监狱服刑人员血样中检测出HIV抗体阳性25例,阳性率为0.58%;在10 587英德监狱的服刑人员血样中检测出HIV抗体阳性36例,阳性率为0.34%。结论 应加大对监狱服刑人员的疾病监测和防病知识宣传力度,降低艾滋病等疾病在监狱服刑人员中及返社会后传播的危险性。     

Myocarditis: an expected health hazard associated with water resources contaminated with Coxsackie viruses type B

Enteroviruses, especially Coxsackie B viruses (CBVs), are responsible for approximately 50% of cases of viral myocarditis. In the present study, serum samples (160) were collected from acute myocarditis patients at different age groups and 104 samples of the same age groups as a control. Cholesterol, LDH, CPK, and GOT were measured for all serum samples (264). Also, to study the source of virus transmission, 72 water and 72 wastewater samples were collected from water and wastewater treatment plants at intakes and outlets. Water and wastewater samples were concentrated by filtration through Zeta-plus filter cartridges and reconcentrated by the PEG-6000 precipitation method. Serum, water, and wastewater samples were inoculated in BGM cells for three successive passages. RT-PCR with enterovirus primers was carried out directly for serum samples and for 1st and 3rd cell culture passages. The positive samples were used for neutralization assay using anti-CBV sera pool to determine the CBV followed by neutralization with separate antisera. The results showed that 50 (31.25%) serum samples from acute myocarditis patients and two (1.4%) samples from the controls were positive for enterovirus RT-PCR. For water and wastewater samples enteroviruses were present in 63.8% and 8.3% for intake and outlet of water treatment plants and, 66.6% and 47.2% for intake and outlet of wastewater treatment plants, respectively. The level of CBV serotypes was varied where CBV3 was dominant for all age groups of myocarditis patients and CBV2 and CBV5 were also detected while CBV2 was the main CBV in water samples and CBV2, 3 and 5 were detected in wastewater samples. The integration of cell culture-PCR reduces the time required for virus detection and enhances the sensitivity of the test.     

Exposure of perfluorinated chemicals through lactation: levels of matched human milk and serum and a temporal trend, 1996-2004, in Sweden

BACKGROUND: Only limited data exist on lactation as an exposure source of persistent perfluorinated chemicals (PFCs) for children. OBJECTIVES: We studied occurrence and levels of PFCs in human milk in relation to maternal serum together with the temporal trend in milk levels between 1996 and 2004 in Sweden. Matched, individual human milk and serum samples from 12 primiparous women in Sweden were analyzed together with composite milk samples (25-90 women/year) from 1996 to 2004. RESULTS: Eight PFCs were detected in the serum samples, and five of them were also above the detection limits in the milk samples. Perfluorooctanesulfonate (PFOS) and perfluorohexanesulfonate (PFHxS) were detected in all milk samples at mean concentrations of 0.201 ng/mL and 0.085 ng/mL, respectively. Perfluorooctanesulfonamide (PFOSA), perfluorooctanoic acid (PFOA), and perfluorononanoic acid (PFNA) were detected less frequently. DISCUSSION: The total PFC concentration in maternal serum was 32 ng/mL, and the corresponding milk concentration was 0.34 ng/mL. The PFOS milk level was on average 1% of the corresponding serum level. There was a strong association between increasing serum concentration and increasing milk concentration for PFOS (r(2) = 0.7) and PFHxS (r(2) = 0.8). PFOS and PFHxS levels in composite milk samples were relatively unchanged between 1996 and 2004, with a total variation of 20 and 32% coefficient of variation, respectively. CONCLUSION: The calculated total amount of PFCs transferred by lactation to a breast-fed infant in this study was approximately 200 ng/day. Lactation is a considerable source of exposure for infants, and reference concentrations for hazard assessments are needed.     

Transport and temperature effects on measurement of serum and plasma potassium

Transport of blood samples from general practice to a central laboratory can result in spuriously high or low potassium concentrations. The importance of this phenomenon was studied in a general practice serving a population of 15,000 patients, 27 km from the pathology laboratory that routinely measured serum potassium. The design involved comparison of potassium levels between control serum (plain gel-separation serum tubes centrifuged in the surgery), routine serum (plain gel-separation tubes centrifuged in the laboratory) and routine plasma samples (lithium-heparin tubes centrifuged in the laboratory). Complete triple sets of data were obtained for 371 samples. Altman and Bland plots for the control serum vs routine serum samples showed a mean difference of +0.1 mmol/L with limits of agreement (+/- 2SD) +0.6 mmol/L, -0.4 mmol/L and for control serum vs routine plasma a mean difference of +0.2 mmol/L with limits of agreement +0.8 mmol/L, -0.4 mmol/L. There was a negative association between mean weekly routine plasma potassium levels with mean weekly temperatures achieved. Regression analysis indicated that both maximum temperature achieved and time to centrifugation significantly contributed to differences observed in the routine plasma samples, but not with the routine serum samples. For plasma samples exposed to high temperatures a clinically significant lowering of potassium concentrations can arise. These results confirm that spurious lowering of potassium concentrations occurs in plasma samples collected in a primary care setting. The preferred method is to centrifuge samples soon after venepuncture. Where this is not possible, collection into plain gel-separation tubes (serum) ensures less variation due to temperature and time to centrifugation than does collection into lithium-heparin tubes (plasma).     

Amoeba-resisting bacteria and ventilator-associated pneumonia

To evaluate the role of amoeba-associated bacteria as agents of ventilator-associated pneumonia (VAP), we tested the water from an intensive care unit (ICU) every week for 6 months for such bacteria isolates; serum samples and bronchoalveolar lavage samples (BAL) were also obtained from 30 ICU patients. BAL samples were examined for amoeba-associated bacteria DNA by suicide-polymerase chain reaction, and serum samples were tested against ICU amoeba-associated bacteria. A total of 310 amoeba-associated bacteria from 10 species were isolated. Twelve of 30 serum samples seroconverted to one amoeba-associated bacterium isolated in the ICU, mainly Legionella anisa and Bosea massiliensis, the most common isolates from water (p=0.021). Amoeba-associated bacteria DNA was detected in BAL samples from two patients whose samples later seroconverted. Seroconversion was significantly associated with VAP and systemic inflammatory response syndrome, especially in patients for whom no etiologic agent was found by usual microbiologic investigations. Amoeba-associated bacteria might be a cause of VAP in ICUs, especially when microbiologic investigations are negative.     

Dried venous blood samples for the detection and quantification of measles IgG using a commercial enzyme immunoassay

OBJECTIVES: To determine whether samples of dried venous blood (DVB) were an acceptable alternative to serum for detecting measles-specific IgG in a commercial enzyme immunoassay. METHODS: Paired samples of serum and DVB were collected from 98 suspected cases of measles and 1153 schoolchildren in Victoria, Australia. All samples were tested using the Dade Behring Enzygnost Anti-Measles-Virus/IgG immunoassay. DVB samples were eluted using either the sample buffer provided with the kit or 5% dry milk powder in phosphate-buffered saline-Tween 20. FINDINGS: DVB samples eluted by sample buffer showed significantly better linear correlation to the serum samples than did DVB samples eluted in 5% dry milk in phosphate-buffered saline-Tween 20. To improve the comparability of serum and DVB samples an adjustment factor of 1.28 was applied to the optical density (OD) values of DVB. This adjustment also enabled quantification of the titre of measles IgG in mIU/ml directly from the OD value using the alpha calculation as specified by the kit protocol. For DVB samples stored for less than six months at 4 degrees C, the assay showed an overall sensitivity of 98.4% and a specificity of 97.2% compared with the results of serum testing. CONCLUSION: These results illustrate the potential for DVB samples to be widely used with the Dade Behring enzyme immunoassay system for determining the immunity of the individual and the population to the measles virus.     

Detection of Zika Virus in Urine

We describe the kinetics of Zika virus (ZIKV) detection in serum and urine samples of 6 patients. Urine samples were positive for ZIKV >10 days after onset of disease, which was a notably longer period than for serum samples. This finding supports the conclusion that urine samples are useful for diagnosis of ZIKV infections.     

The effect of timing of sample collection on the detection of measles-specific IgM in serum and oral fluid samples after primary measles vaccination

This study compares the timing of the rise and decline of measles-specific IgM in serum samples and in oral fluid samples. Two hundred and eighty 9-month-old infants presenting for routine measles vaccination in Addis Ababa, Ethiopia, were enrolled. Paired serum and oral fluid samples were collected before and 1, 2, 3 or 4 weeks after measles vaccination. Samples were tested by using a modified antibody-capture enzyme immunoassay. For the 321 IgM-negative pre- and post-vaccination serum samples, 317 (99 %) of their corresponding oral fluid samples were IgM-negative. Among the 130 IgM-positive serum samples, 75% of their paired oral fluid samples were IgM-positive, with the percentage rising to 87% after oral fluid samples collected > or =3.5 weeks after vaccination were excluded. Among the post-vaccination serum samples, the percent IgM-positive peaked in week 3 and declined to 79% in week 4. For post-vaccination oral fluid samples, the percent IgM-positive peaked in weeks 2 and 3, and then declined to 43% in week 4. This modified antibody-capture enzyme immunoassay appears to detect vaccine-induced measles-specific IgM in the first 3 weeks after vaccination.