Naja melanoleuca cobra venom contains two forms of complement-depleting factor (CVF)

Two forms of complement-depleting cobra venom factor (CVFm1 and CVFm2), possessing molecular masses of 142.6 kDa (CVFm1) and 143.1 kDa (CVFm2), according to MALDI mass-spectrometry, were isolated from the Naja melanoleuca cobra venom. As shown by polyacrylamide gel electrophoresis in the presence of SDS, both forms similarly to factor from the Naja kaouthia cobra venom (CVFk) consist of three polypeptide chains with molecular masses of about 70, 50, and 30 kDa, the two large subunits being glycosylated. As determined by MALDI mass-spectrometry, 30 kDa subunits of CVFm1 and CVFm2 have considerably different finger-prints of tryptic digests that suggests differences in their amino acid sequences. A study of activity in vivo has shown no significant differences in C3 consumption by CVFm1, CVFm2 and CVFk in mouse blood. However, as shown by an immunoassay method, they differ in their ability to activate the complement system via C3 conversion, the ratio of these activities for CVFm1:CVFm2:CVFk being 2.5:1.6:1. Kinetic studies using a hemolytic test showed that complement depletion by CVFm1 is faster than that by CVFm2. Thus, for the first time the presence in a single venom of two forms of CVF differing by both amino acid sequence and biological activity has been shown.     

医务人员与SARS患者免疫指标的对比研究

目的 :研究严重急性呼吸道综合征 (SARS)病毒对医务人员免疫功能可能产生的影响 ,为传染病的防治提供理论依据。方法 :采用胶体金法和ELISA法检测SARS患者 (SARS患者组 )及与患者密切接触的医务人员 (小汤山医院组与非SARS定点医院组 )血清中SARSIgG抗体含量 ,并用特种蛋白分析仪测定CRP、补体C3、C4及CIC含量 ,同时选取 13例正常人作为对照组。结果 :对照组血清SARSIgG抗体均阴性 ;非SARS定点医院组医务人员血清中 ,胶体金法 1例阳性 ,ELISA法 1例阳性 ,阳性率为 1 14 % ;SARS患者组IgG抗体均阳性 ,阳性率 10 0 %。SARS患者组CRP分别是对照组、非SARS定点医院组、小汤山医院组的 3 1倍、3 5倍和 2 8倍(P <0 0 1)。与对照组比较 ,非SARS定点医院组补体C3、C4下降了 2 5 2 0 %和 2 7 5 9% ,小汤山医院组补体C3下降了 13 3 9% (P <0 0 5 )。非SARS定点医院组补体C3、C4与小汤山医院组比较 ,分别下降了 13 64 %和 2 2 2 2 % (P <0 0 5 ) ,CIC含量增高了 2 0 0 2 % (P<0 0 5 )。结论 :SARS可能存在隐性感染 ,CRP与患者的临床症状相关 ,可作为SARS病情监测的指标 ;而病毒对医务人员免疫功能的影响显示 ,小汤山医院组小于非SARS定点医院组 ,表明小汤山医院的隔离防护体系较为完善     

产前检查免疫检验项目应用的价值评估

目的分析产前检查免疫检验项目的应用价值。方法选取2016年11月～2018年1月在我院进行分娩的产妇80例作为观察对象,将80例产妇分成免疫组与常规组,各组40例。常规组产前实施常规检验,免疫组则在常规检验的基础之上进行免疫项目检验,观察两组方法的临床检验效果。结果免疫组孕妇乙型肝炎、梅毒与微生物感染发生率和高危妊娠发生率要显著低于常规组孕妇,差异有统计学意义(P0.05)。80例孕妇的HIV抗体、乙肝抗体、丙肝抗体等阳性检出率分别为0(0/80)、7.5%(6/80)、5.0%(4/80)。结论孕妇在产前进行免疫项目检验,可以早期对孕妇身体状况与胎儿的健康状态实施检测判断,能够确保母亲与婴儿的安全,降低产妇妊娠与分娩时期的并发症发生率,为产妇安全分娩提供重要保障。     

Continuous reference intervals for 19 endocrine,fertility, and immunochemical markers in the CALIPER cohort of healthy children and adolescents

BackgroundReference intervals are traditionally partitioned into discrete ranges by major covariates such as age and sex. However, discrete reference intervals often oversimplify the complex relationship between analyte concentration and age. Continuous reference intervals have been suggested to more accurately represent this complex relationship, particularly in pediatrics. The objective of this study was to establish continuous reference intervals for endocrine, fertility, and additional immunochemical parameters in the CALIPER cohort of healthy children and adolescents.MethodsContinuous reference intervals from 1 to 18.5 years of age were established using retrospective CALIPER data collected from healthy Canadian children and adolescents. Continuous reference intervals (2.5th and 97.5th percentiles) were determined for 19 parameters by nonparametric quantile regression. Total and yearly flagging rates were calculated for the upper and lower continuous reference limits and compared to previously published partitioned reference limits.ResultsContinuous reference intervals were established for 19 endocrine, fertility, and additional immunochemical parameters, with 11 requiring sex-specific reference curves. Continuous reference intervals assessed both visually and by flagging rate analysis more accurately represented the relationship between analyte concentration and age, particularly for parameters with complex reference value patterns.ConclusionThis is the first comprehensive report to establish continuous reference intervals for several immunochemical parameters including endocrine and fertility markers in a healthy paediatric Canadian cohort. The ability of continuous reference intervals to provide a better estimate of age-related changes in reference values suggest their potential to improve paediatric laboratory test result interpretation and clinical decision-making.     

Measurement Of Angiotensinogen In Human Serum By Fluorescence Polarization Immunoassay

Fluorescence polarization immunoassay (F.P.I.A.) has rarely been used to measure components of the renin system. Using an Abbott TDX polarimeter and fluorescein - labelled angiotensin I as a tracer we measured angiotensin I by F.P.I.A. Combining this procedure with a renin incubation step enabled measurement of angiotensinogen in human serum. Using sera from male patients and from pregnant females, a good correlation between radioimmunoassay (R.I.A.). and F.P.I.A. was found. Two procedures were developed; one involving taking samples from the renin incubation mixture, the other involving measurement of generated angiotensin I at intervals without interrupting the renin incubation procedure. F.P.I.A. is less expensive and somewhat simpler than R.I.A. but, with the instrument used, it     

Evaluation of commercial multi-drug oral fluid devices to identify 39 new amphetamine-designer drugs

Recently, the diffusion on the black market of new psychoactive substances not controlled and often sold as ‘legal highs’, is exponentially increasing in Europe. Generally, the first analysis for these drugs involves an immunoassay screening in urine or plasma. Actually, there is growing interest in the use of oral fluid (OF) as alternative specimen over conventional biological fluids for drug testing, because of the significant advantages, as a non-invasive collection under direct observation without undue embarrassment or invasion of privacy, and a good correlation with plasma analytical data. Few assays have been developed for detection of new psychoactive compounds in biological samples, so it is important to investigate how they may or may not react in pre-existing commercial immunoassays. In this paper, two different multi-drugs oral fluid screen devices (OFDs) (Screen® Multi-Drug OFD and GIMA One Step Multi-Line Screen Test OFD) were evaluated to determine the cross-reactivity of thirty-nine new amphetamine designer drugs, including twelve substances officially recognized as illicit by italian legislation. Cross-reactivity towards most drugs analyzed was <1 in assays targeting amphetamine (AMP) or methamphetamine (MET). Only two (p-methoxyamphetamine and p-methoxymethamphetamine) of all tested amphetamines gave a positive result.     

Vitellogenin, steroid plasma levels and spawning performance of cultured female Senegalese sole (Solea senegalensis)

The Senegalese sole (Solea senegalensis) is a high value market flatfish, which aquaculture is compromised by severe reproductive problems; these are mostly found in soles hatched and raised in captivity (F1 generation). To gain knowledge on the reproductive dysfunctions observed in cultured (F1) Senegalese sole, this work aimed at developing a specific vitellogenin (VTG) ELISA, for the measurement of plasma VTG levels in this species. Profiles of VTG were correlated with those of sexual steroids and spawning performance of an F1 broodstock, during three consecutive years. The Senegalese sole VTG (ssVTG) was purified by precipitation with MgCl(2)-EDTA and anion-exchange chromatography and showed a molecular mass of 172 kDa, by SDS-PAGE. Specific antibodies were obtained and used to develop a competitive ELISA, which had a sensitivity of 3.6 ng ml(-1), and inter- and intra-assay coefficients of variation of 9.5% (n=29) and 6.7% (n=12), respectively. Annual profiles of plasma VTG showed a major peak at pre-spawning, and a second minor rise around autumn, which mirrored plasma profiles of both estradiol (E(2)) and testosterone (T) levels. Spontaneous spawning occurred every year in the spring season, but no fertilized eggs were obtained. In conclusion, this study described, for the first time, the purification and development of a sensitive and specific ELISA for Senegalese sole VTG. The endocrine and spawning data suggested that F1 female broodstock showed normal VTG and steroid releasing profiles in captivity with occurrence of spontaneous spawning, but no fertilization of the eggs was recorded.     

快速免疫色谱测定法在骨关节结核诊断及鉴别诊断中的价值

目的探讨快速免疫色谱法对活动性骨关节结核的诊断价值。方法采用ＩＣＴ快速免疫色谱法检测７５例活动性骨关节结核、６９例以肿瘤为主的骨病患者及７２名正常人血清中的抗体。结果骨关节结核组阳性率为８０％，其中脊柱结核阳性率为９０％，脊柱以外的关节结核阳性率为６１％，合并其它部位结核阳性率为７６％；骨病组３例阳性，正常组２例阳性，假阳性率分别为４％和３％。结论ＩＣＴ快速免疫色谱法是骨关节结核诊断和鉴别诊断的一项有用的辅助手段     

TIME-RESOLVED IMMUNOFLUOROMETRY OF SERUM HTSH WITH ENHANCED SENSITIVITY

ABSTRACT

Sensitive TSH immunoassays offer a clear advance in discriminating the TSH concentrations in serum between hyperthyroid and euthyroid individuals; they have been proposed as the best single screening test for thyroid disorders. We have developed a highly sensitive serum TSH TRFIA based on DELFIA technology. Three monoclonal antibodies (McAbs) directed against different epitopes of the TSH molecule were involved in this assay, of which, one McAb was used to coat clear microwells, and the other two were biotinylated for signal generation after being bound by the europium labeled streptavidin. The europium label captured on the well surface was quantified by a routine dissociation-enhancement procedure. The fluorescence intensity was directly proportional to the serum hTSH concentration. The assay required two steps and could be completed within 5 h. The analytical sensitivity reached 0.002 mIU/L with a sample volume of 100 µL, the function sensitivity was 0.017 mIU/L. Measurements by the present method correlated well with that obtained by the ACS-180 chemi-luminescence immunoassay (CLIA). The discrimination of hyperthyroid patients from clinically euthyroid patients by the present method was much better than that by the routine IRMA.