Counterimmunoelectrophoresis compared with complement fixation and passive haemagglutination tests in the evaluation of the immune response in Campylobacter infections

The immune response was studied in 238 human patients with Campylobacter jejuni/coli (CJC)-infections in Rotterdam by the counterimmunoelectrophoresis (CIE) test, a commercial complement fixation test (CFT) and the passive haemagglutination test (HA). Antibodies became detectable in the three tests around 4 days after the onset of complaints. Between the 7th and the 20th days after onset of illness 79%, 80% and 53% of the patients demonstrated antibodies by the CIE, the CFT and the HA, respectively. The HA took 30 days to reach 60% positive serum samples and this percentage declined to 35 by the 50th day. Antibody titres demonstrated in the CIE and the CFT declined more slowly. CIE and CFT performed with antigens from a limited number of heat-stable serotypes can be used in the evaluation of the humoral immune response in CJC-infections.     

Glomerulonephritis Associated with Antibody to Glomerular Basement Membrane

Glomerulonephritis associated with antibody to glomerular basement membrane, shown by linear staining of the glomerular basement membrane with fluoresceinated anti-IgG antisera, was found in only 10 out of 400 (2·5%) renal biopsy specimens studied by immunofluorescence. Seven of these cases had rapidly progressive glomerulonephritis, five with lung haemorrhage (Goodpasture''s syndrome) and two without, and three had less severe nephritis without lung haemorrhage. Circulating antibody to glomerular basement membrane, measured by a passive haemagglutination technique and by indirect immunofluorescence, was detected in the serum of all patients with rapidly progressive glomerulonephritis by both techniques but only by the passive haemagglutination method in two of the other three patients. Two patients died of their lung haemorrhage, one despite bilateral nephrectomy, and lung haemorrhage and circulating antibody to glomerular basement membrane persisted after bilateral nephrectomy in another patient.     

Etiology of acute and chronic bronchopulmonary diseases in children. II. A study of the specific immune response by counterimmunoelectrophoresis and the complement fixation and passive hemagglutination reactions

Specific immune response to Streptococcus pneumoniae and Haemophilus influenzae has been studied in 158 children with acute pneumonia and pleuritis and 128 children with chronic pneumonia by countercurrent immunoelectrophoresis (CIE) and in the complement fixation (CFT) and passive hemagglutination (PHA) tests. The use of CIE leads to the detection of antibodies to H. influenzae in 23.7% of children with acute pneumonia and in 46.9% of children with chronic pneumonia. In the CFT antibodies to H. influenzae are also more often detected in children with chronic pneumonia (48%) than in those with acute respiratory infections (12.2%). In the PHA test high titers of antibodies to type b H. influenzae capsular polysaccharide occur in 11.9% of children with acute pneumonia and in 8.2% of children with chronic pneumonia.     

HBs antigenemia in glomerulonephritis in children

Altogether 194 glomerulonephritis patients were examined by three methods: countercurrent immunoelectrophoresis, passive hemagglutination test, and enzyme immunoassay. Use of the most sensitive method, viz. enzyme immunoassay, has yielded the highest HBsAg detection rate: 29.1% in acute glomerulonephritis and 21.4% in chronic glomerulonephritis. This method may be recommended for the examination of glomerulonephritis patients whose sera contain HBsAg in low titers.     

Comparison of enzyme-linked immunosorbent assay, electron microscopy, and reversed passive haemagglutination for detection of human rotavirus in stool specimens

An enzyme-linked immunosorbent assay (ELISA) using microplates as solid phase, rabbit antiserum against human rotavirus Wa strain as catching antibody, and the same reagent labeled with beta-D-galactosidase as conjugate, has been developed for detection of human rotavirus antigen(s) in stool specimens from patients with acute gastroenteritis. The limit of detection of purified human rotavirus by ELISA was 15.6 ng/ml (1.56 ng/well) of viral protein. The sensitivities of ELISA, electron microscopy, and the reversed passive haemagglutination method (ROTA-CELL) were compared. ELISA was more sensitive than electron microscopy and the reversed passive haemagglutination method. The ELISA blocking assay was useful for detection of an antibody response to human rotavirus in paired sera from children in two institutions during outbreaks of rotavirus gastroenteritis.     

Changes in collagen metabolism--another look at osteoarthrosis

Collagen types I and III were isolated from osteoarthrotic cartilage. Immunofluorescence study has shown that these two collagen types are present in the deep layers of cartilage. Additional staining for fibronectin revealed the presence of the cell-adhesive protein in osteoarthrotic cartilage. Neither collagen type I and type III nor fibronectin were found in control cartilage. A passive haemagglutination assay determined anticollagen antibodies (against types I, II and III) in sera of some osteoarthrotic patients.     

Enzyme-linked immunosorbent assay (ELISA) for IgM, IgG and IgA class antibodies against Candida albicans antigens: development and comparison with other methods

An extract of Candida albicans was used as an antigen on microtitre plates in the enzyme-linked immunosorbent assay (ELISA) to measure IgM, IgG and IgA class antibodies in the sera of hospitalized patients. It was found that of these patient sera that reacted positively in Ouchterlony immunodiffusion (ID) when undiluted, 58% were also positive in the ELISA against the same antigen preparation. However, all the sera with an ID titre of 1:2 or higher were ELISA-positive, demonstrating especially IgG and IgA. Of the sera positive by counterimmunoelectrophoresis against somatic and metabolic antigens of C. albicans, 86% were positive by ELISA. Reactions in precipitin-negative sera, if they occurred, usually demonstrated IgM or IgA. The sera with high passive haemagglutination or indirect immunofluorescence titres against surface antigens of C. albicans were positive in the IgG and IgA assays, while approximately one third were positive in the IgM assay.     

A chimera antibody erythroimmunoassay for detecting HBsAg in human sera

A highly efficient chimera antibody, a monoclonal anti-hepatitis-B-surface (anti-HBs) antibody coupled with polyclonal anti-sheep-red-blood-cell (anti-SRBC) antibody was prepared using a heterobifunctional reagent, N-succinimidyl-3-(2-pyridyldithiopropionate) (SPDP). Using SRBC as a marker, we established a sensitive solid-phase chimera antibody erythroimmunoassay (CAEIA) according to Guesdon's method. The sensitivity of this assay was 2–20 times higher than the reverse passive haemagglutination assay (RPHA) for detecting HBsAg in serially diluted sera from 10 hepatitis B patients. The weakest quantity of HBsAg detected by this assay was 4.5 ng/ml, while RPHA was unable to detect less than 75 ng/ml of HBsAg. The assay was as sensitive as the enzyme-linked immunosorbent assay (ELISA) and gave more accurate, reproducible and stable results than ELISA; specificity was also satisfactory.     

Evaluation of three serological tests for the detection of human plague in northeast Brazil.

The passive haemagglutination (PHA) test, enzyme-linked immunosorbent assay (ELISA) and the dot enzyme-immunosorbent assay (DOT-ELISA) were used to detect the levels of IgG antibodies against the Fraction 1 (F1) antigen of Yersinia pestis in sera of plague-infected patients from Northeast Brazil. Twenty three selected PHA-positive sera of subjects with bacteriological confirmation of plague were also positive in the DOT-ELISA but only 19 were detected by the conventional ELISA technique. Another group of 186 serum samples from subjects diagnosed as plague-infected by clinical and epidemiological parameters, but PHA-negative, were screened with DOT-ELISA and 11 gave positive results. The specificity of the assays on the serological detection of plague was confirmed in inhibition tests using purified F1 antigen. These results suggest that DOT-ELISA can be an useful, simple and more sensitive alternative for the serodiagnosis of plague in Northeast Brazil.     

A study of factors affecting the sensitivity of the passive haemagglutination method for serotyping Campylobacter jejuni and Campylobacter coli and recommendations for a more rapid procedure

Factors affecting the sensitivity of the passive haemagglutination method for serotyping campylobacters have been studied. The concentration of red blood cells during the haemagglutination stage of the procedure markedly affected the titer obtained. An increase in concentration of red blood cells resulted in a lower titer, with titers being inversely proportional to red blood cell concentration. No differences in titer were observed when erythrocytes were sensitized at a range of pH values between pH 5.0 and pH 8.0. The time required for antigen extraction and for red blood cell sensitization was shown to be 15 min each, thus resulting in a reduction in the time required for serotyping. Furthermore, use of avian erythrocytes enabled the haemagglutination reactions to be read after incubation for only 1 h. Combining these procedures with a rapid slide haemagglutination test enables a single worker to serotype over 100 C. jejuni and C. coli isolates within 1 working day.     

Detection and titration of antitetanus antibodies using an automated passive hemagglutination method]

An automated reversed passive haemagglutination technique is described, using human erythrocytes sensitized by the chromic chloride method with higly purified tetanus toxoid. Such erythrocytes are agglutinated by the specific antibodies in a Technicon autoanalyzer. Clots are decanted, supernatant is hemolyzed and optical density is measured at 550 millimicron. Assay standards for comparison, ranging from 5 to 25 UI/ml, are prepared from human antitetanus immunoglobulins titrated by toxin neutralisation assay in mice. This method allows to screen donors' sera for presence of high titers of tetanus antibodies (larger than or equal to 5 UI) suitable for preparation of antitetanus immunoglobulins. 4,4% of donors have circulating antitetanus antibody levels corresponding to 5 UI/ml or more, by this method. The specificity of antibodies has been confirmed by the neutralisation assay in mice. The results obtained among 1.000 donors well agree with those of the counter-immuno-electrophoresis technique (C.E.P.). But, when, compared with C.E.P., the haemagglutination assay appears more objective, more quantitative and sensitive, and allows to get not only a rapid screening test but also a precise titration simultaneously.     

Comparison of five tests for the serologic diagnosis of myiasis by Gasterophilus spp. larvae (Diptera: Gasterophilidae) in horses and donkeys: a preliminary study

Abstract. Sera from 41 horses and 159 donkeys, from twelve States of México, were tested to ascertain anti-Gasterophilus circulating antibodies by double immunodiffusion (DD), counterimmunoelectrophoresis (CIE), indirect haemagglutination (IH), thin layer immunoassay (TIA) and diffusion-in-gel ELISA (DIG-ELISA) methods using crude somatic antigen from third instar larvae of G.intestinalis (DeGeer). At necropsy, 33/41 horses and 24/159 donkeys were found to be parasitized by G.intestinalis and/or G.nasalis (L.). Gasterophilus intestinalis was the species most commonly found in the equines. Analysis of the sera from the infested animals by DD showed positive results of 21.2% in horses and of 8% in donkeys. Screening the sera with CIE gave sensitivities of 69.7% in horses and of 32% in donkeys. Examination of the sera by IH showed positive results of 87.9% and of 48% in horses and donkeys, respectively. Testing the sera with TIA gave sensitivities of 93.9% in horses and of 96% in donkeys. Analysis of horses' sera by DIG-ELISA showed a sensitivity of 93.9%.     

Usefulness of the passive hemagglutination micromethod in the diagnosis of whooping cough

Studies on modification of diagnostic test for pertussis have been continued, they were practically restricted to an application of passive haemagglutination micromethod. Six hundred and twenty eight sera from children suspected to be infected with Bordetella and 38 sera of control children suffering from non-infectious diseases were tested. Despite of the fact that higher titers were obtained with method using red blood cells preserved in Alsevier solution, the passive haemagglutination micromethod may be used in field studies especially during pertussis epidemics what was confirmed by statistical analysis.     

慢性肾小球肾炎患者血清HGF、Cys-C、TAFI水平变化及其临床诊断价值

目的:分析慢性肾小球肾炎患者血清肝细胞生长因子(HGF)、胱抑素C(Cys-C)、凝血酶激活的纤溶抑制物(TAFI)水平的变化及其临床诊断价值。方法:选择我院2017年1月～2018年5月收治的71例慢性肾小球肾炎患者作为慢性肾小球肾炎组及同期于本院进行健康体检的83例作为健康对照组。检测进而比较两组血清HGF、Cys-C、TAFI水平,分析以上指标和患者肾功能的相关性及对慢性肾小球肾炎的诊断价值。结果:慢性肾小球肾炎组血清HGF、Cys-C、TAFI水平均显著高于对照组(P0.05)。慢性肾小球肾炎患者治疗后血清HGF、Cys-C、TAFI水平均显著低于治疗前(P0.05)。慢性肾小球肾炎患者血清HGF、Cys-C、TAFI水平和肾功能指标(肌酐(Scr)、尿素氮(BUN)、尿酸(UA))均呈显著正相关(P均0.05)。血清HGF水平诊断慢性肾小球肾炎的曲线下面积为0.826,敏感度和特异度分别为0.747和0.746;血清Cys-C水平诊断慢性肾小球肾炎的曲线下面积为0.821,敏感度和特异度分别为0.687和0.859;血清TAFI水平诊断慢性肾小球肾炎的曲线下面积为0.816,敏感度和特异度分别为0.855和0.647;血清HGF、Cys-C、TAFI水平联合检测诊断慢性肾小球肾炎的曲线下面积为0.951,敏感度和特异度分别为0.831和0.757。结论:慢性肾小球肾炎患者血清HGF、Cys-C及TAFI水平均明显升高,联合检测血清HGF、Cys-C及TAFI可能作为慢性肾小球肾炎诊断及预评估参考指标。     

Characterization of strain specific typing antisera for genetic monitoring of inbred strains of rats

Strain-specific typing antisera (SSTA) were prepared for six inbred strains of rats by using a pooled immunization protocol. The SSTA were used in both a haemagglutination assay and a complement dependent microcytotoxicity assay to compare the usefulness of the two test systems. Both assays were simple, reliable and repeatable, and each system had distinct advantages and disadvantages. The haemagglutination assay was faster and required less specialized equipment than the microcytotoxicity assay. On the other hand, interpretation of results in the microcytotoxicity assay was easier and more objective. It was concluded that SSTA could be used with the microcytotoxicity assay and/or the haemagglutination assay to provide a simple and effective genetic monitoring method for inbred strain of rats.     

The persistence of antibodies induced by meningococcal polysaccharides of groups A and C in human volunteers

Ninety-nine college students (58 males and 41 females) aged 17 to 23 years were each injected subcutaneously with 50 micrograms of meningococcal polysaccharides A and C. Titrations of antisera obtained at various time intervals during the two subsequent years were made by passive haemagglutination microtitration using human O Rh-negative red blood cells from a single source. The percentage of responders (those who developed a fourfold increase in titre) to polysaccharide A was 97.9% and that to polysaccharide C was 94.8%. Pre-immunization titres of 16 to polysaccharide A and 8 to polysaccharide C were considered to be threshold values above which the response might be impaired. There was a significant (P less than 0.01) difference in the geometric mean HA titre between vaccinees and control groups after vaccination at each time interval studied. The majorities of both the vaccinees and the controls had a higher peak titre to polysaccharide A than to polysaccharide C.     

Detection and titration of anti-tetanus antibodies by passive hemagglutination]

Human red cells O Rh + are coated with purified tetanus toxoid, by means of chromic chloride as coupling agent. With this reagent and by passive haemagglutination technique, antitetanus antibodies can be automatically screened and titrated in blood donnors plasmas, on Groupamatic equipments. Comparative results of three techniques: counter immunoelectrophoresis; manual passive haemagglutination; automatic passive haemagglutination are given, 4, 92 percent among the 1.219 tested blood donnors plasmas have an antibody greater than or equal to 5 U.I., 16, 24 percent have a titre between 1 and 5 U.I.     

Comparative serology techniques for the diagnosis of Trypanosoma cruzi infection in a rural population from the state of Querétaro, Mexico

Immunological diagnostic methods for Trypanosoma cruzi depend specifically on the presence of antibodies and parasitological methods lack sensitivity during the chronic and “indeterminate” stages of the disease. This study performed a serological survey of 1,033 subjects from 52 rural communities in 12 of the 18 municipalities in the state of Querétaro, Mexico. We detected anti-T. cruzi antibodies using the following tests: indirect haemagglutination assay (IHA), indirect immunofluorescence assay (IFA), ELISA and recombinant ELISA (rELISA). We also performed Western blot (WB) analysis using iron superoxide dismutase (FeSOD), a detoxifying enzyme excreted by the parasite, as the antigen. Positive test results were distributed as follows: ELISA 8%, rELISA 6.2%, IFA and IHA 5.4% in both cases and FeSOD 8%. A comparative study of the five tests was undertaken. Sensitivity levels, specificity, positive and negative predictive values, concordance percentage and kappa index were considered. Living with animals, trips to other communities, gender, age, type of housing and symptomatology at the time of the survey were statistically analysed using SPSS software v.11.5. Detection of the FeSOD enzyme that was secreted by the parasite and used as an antigenic fraction in WBs showed a 100% correlation with traditional ELISA tests.     

Frequency of human toxocariasis in Jos, Plateau State, Nigeria

The enzyme-linked immunosorbent assay (Elisa) was used to examine sera of 104 children and adults in Jos, Plateau State, Nigeria for anti-toxocaral antibodies, out of which 31 (29.8%) were reactive. The seropositive rates were 30.4% for adults, 29.6% for children, 34% for females and 25.9% for males. However, the differences were not significant by age and sex. A highly significant association (p < 0.001) was observed between seropositivity and geography but none between seropositivity and dog ownership (p > 0.05).     

Chronic renal failure in children.

Seventy-seven children with chronic renal failure were examined at one hospital in the province of Quebec between 1970 and 1975; this represents an incidence of 2.5 per million population per year. The entities responsible for chronic renal failure were urinary tract malformation (in 36%), chronic glomerulonephritis (in 22%), congenital renal parenchymal malformation (in 21%) and hereditary nephropathy (in 13%). The evolution of chronic renal failure in children with either vesicoureteral reflux or a posterior urethral valve seemed to be related more to the initial severity of the disease than to the age at the time of diagnosis. Hence any screening program designed to detect kidney disease in schoolchildren would not prevent chronic renal failure, since at that age renal parenchymal damage seems to be irreversible. The manner in which chronic glomerulonephritis evolved depended on whether the nephrotic syndrome was present and on the type of histologic lesion. Children with congenital renal hypoplasia or dysplasia often presented with seizures due to hypertensive encephalopathy without obvious symptoms or signs of pre-existing renal disease. Among patients with familial nephropathy many of those with cystinosis underwent successful renal transplantation early in life.