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At least 109 records · Page 6Linked to original sources
PubMed2005

[ELISA study on application of recombinant glycoprotein D of herpes simplex virus-1 in diagnosis of herpes simplex virus-1 infection with ELISA].

OBJECTIVE: To establish a specific and sensitive serological ELISA, diagnostic method, for herpes simplex virus-1 (HSV-1) infection using yeast expressed glycoprotein D (gD) of HSV-1 as coating antigen. METHODS: The yeast expressed products were collected at the most appropriate time and sonicated. After the optimum dilution titers of the coating antigens and sera were determined, 57 clinical sera were assayed using the recombinant gD and HSV-1-infected culture media as coating antigens respectively. The same sera were also assayed by homemade and Euroimmun ELISA kit, Germany. The results of German kit as a golden standard were compared with those of the other three methods in specificity, sensitivity and accordance rate. RESULTS: Compared to the imported kit, the specificities of recombinant protein, HSV-I culture media and homemade kit were 57.1%, 57.1% and 100.0%, respectively, the sensitivities were 82.0%, 78.0%, 48.0% and the accordance rates were 78.9%, 75.4%, 54.4%, respectively. The results of repeated experiments of recombinant protein showed that there was no statistically significant difference between the two experiments (P > 0.05). CONCLUSION: The ELISA with recombinant gD protein of HSV-1 as coating antigen is a specific, sensitive, quick and convenient method for diagnosis of HSV-1 infection.

Animals↗
PubMed2005

Development of the rapid and simple elisa (whole blood-ELISA) using coconut water-tween as a wash solution for whole blood sample from Schistosoma japonicum-infected rabbit and human.

PBS-Tween as a wash solution, prepared with distilled water, is used in ELISA. In areas where schistosomiasis is endemic, however, distilled water is hard to come by. We have modified a WHOLE BLOOD-ELISA test to use coconut water-Tween as a wash solution, because coconut water is easy to come by and cheap in the tropics. We applied the test to whole blood samples from rabbits and humans infected with Schistosoma japonicum. This modified WHOLE BLOOD-ELISA was confirmed to be a rapid, simple, and cost-effective method.

Animals↗
PubMed1988

[Comparison of the IFA, ELISA IgG, ELISA IgM, IHA and the direct agglutination tests in Toxoplasma gondii serology].

In this study, toxoplasma antibodies were evaluated by using IFA, ELISA IgG, ELISA IgM, IHA and direct agglutination test in serum samples obtained from reproductive women. The specificity, sensitivity, unit costing and timing of tests compared with IFA test. In this evaluation IFA test are used. ELISA IgG specificity 98%, sensitivity 97.5%; IHA specificity 90%, sensitivity 97.5%; direct agglutination specificity 92%, sensitivity 95% have been found.

Agglutination Tests↗
PubMed1993

[Serodiagnosis of Pseudomonas aeruginosa infections in mucoviscidosis: comparative study of Western blotting, ELISA exotoxin A and ELISA phospholipase C].

Anti-Pseudomonas aeruginosa antibodies were studied by Western Blot, ELISA-exotoxin A and ELISA-phospholipase C for 91 serums from 31 patients with cystic fibrosis. More, for the two enzyme-linked immunosorbent assays, 44 serums from 44 healthy individuals were studied as controls. The study of these three parameters revealed the followings: with no infection by Pseudomonas aeruginosa all the results were negative, at the beginning of the infection, anti-exotoxin A antibodies appeared in first, followed in some cases by the reactions of Western Blot, anti-phospholipase C antibodies became positive at last and went on a par with the installation of the chronic characteristic of the infection, as soon as the chronicity were indisputable, the three methods revealed elevated serum antibodies amounts. Generally there was a correlation between detected antibodies and Pseudomonas aeruginosa isolation in sputum. Among these three methods, ELISA-exotoxin A appeared to be the most interesting because of its good reproducibility and its early positivity, before the others methods and sometimes before Pseudomonas aeruginosa isolation. It would be a significant argument to establish as soon as possible an antimicrobial therapy.

ADP Ribose Transferases↗
PubMed1993

[Serologic diagnosis of canine leishmaniasis: the dot-ELISA as a simple alternative to conventional ELISA].

Serology is an important tool for the diagnosis of canine leishmaniasis, since the direct detection of the parasite remains negative in many cases. A dot-ELISA was developed, which can be read without using a photometer. For this purpose assay strips were prepared, which were divided into a control field (staining indicates normal function of reagents) and four reaction fields. The reaction fields were coated with four different concentrations of Leishmania antigen and allowed semiquantitative readings of test results. The comparison of the reactions of 73 sera (20 healthy dogs, 24 dogs with leishmaniasis, 15 dogs with trypanosomiasis and 14 dogs with babesiosis) in the dot-ELISA with the results of a conventional microtiter plate ELISA showed a significant correlation between the two methods.

Animals↗
PubMed1986

A comparison of five different methods for the detection of TNP specific mouse IgE: ELISA, ELISA on cells, rosetting, granule enzyme release assay and passive cutaneous anaphylaxis.

Using the same anti-TNP hybridoma supernatant pool as IgE antibody source (2 micrograms/ml), several methods were compared for their sensitivity in IgE detection and convenience for screening purposes. Using rat basophilic leukemia (RBL) cells as specific receptor cells, one out of two rosetting methods with TNP-sheep red blood cells allowed the detection of 0.5 ng IgE/ml (50 pg/assay) while the other was much less sensitive (60 ng/ml). More convenient for screening was an ELISA method performed on cells, in which the IgE bound to the RBL cell surface could be detected either by enzyme-anti-Ig or by enzyme-antigen conjugates with a similar, albeit low, sensitivity of 10 ng IgE/ml (500 pg/assay). Using the same antibody and cell sources, a very convenient and more sensitive method for screening purposes was found to be the measurement of antigen-induced basophil granule beta-N-acetylglucosaminidase release by IgE sensitized RBL cells: 0.5 ng IgE/ml and 50 pg/assay. For the same anti-TNP IgE source, this compares with a detection limit by ELISA of 0.2 ng IgE/ml (10 pg/assay) and by passive cutaneous anaphylaxis in rats of 2 ng IgE/ml (100 pg/assay).

Acetylglucosaminidase↗
PubMed1987

Novel ELISA and ELISA-spot assays used to quantitate B cells and serum antibodies specific for T cell and bromelated mouse red blood cell autoantigens.

The frequency of splenic B cells producing antibodies reactive with bromelain-treated mouse red blood cells (BrMRBC) or T cell surface antigens was examined in autoimmune and normal mice. This was accomplished by fixing target cells to microtiter plates such that their membrane antigens could be detected in ELISA and ELISA-spot assays. This technique was rapid, sensitive, and permitted antibodies of both the IgG and IgM isotypes to be measured independently. Autoimmune NZB, BXSB male and MRL-lpr/lpr mice had 10-100-fold higher levels of serum anti-BrMRBC and anti-T cell antibodies than did control DBA/2 and CBA/J animals. The frequency of splenic B cells producing autoantibodies of these specificities was similarly increased among autoimmune mice. In general, the number of antibody-forming cells (AFC) reactive with BrMRBCs was 2-5 times higher than the number reactive with T cell surface determinants. In NZB mice these cells produced primarily IgM autoantibodies whereas in MRL-lpr/lpr animals they secreted primarily IgG. The concentration of serum autoantibody did not precisely correlate with AFC frequency, indicating that immunoglobulin catabolism and other factors play a role in regulating serum antibody concentration.

Animals↗
PubMed1998

Reduction of singleton reactors against swine vesicular disease virus by a combination of virus neutralisation test, monoclonal antibody-based competitive ELISA and isotype specific ELISA.

Pigs which are serologically positive for swine vesicular disease virus (SVDV) but which show no clinical signs and for which there is neither a relevant history of the disease on the holding nor contact with a known outbreak are considered as singleton reactors. False positive serological results for an epizootic disease, like SVD, in a non-vaccinated population or in imported animals are of great concern to international trade. For the virus neutralisation test, the gold standard for SVD, singleton reactors are found at a level of 1-3/1000. Singleton reactors also occur when other serological testing methods are used. The number of animals finally considered as singleton reactors can be reduced considerably by performing three different serological tests (virus neutralisation test, monoclonal antibody-based competitive ELISA and isotype specific ELISA) on the same serum. A serological profile of the animal can be derived by analysing the results in greater detail. This procedure can reduce considerably the number of pig holdings on which a prohibition of movement and trade needs to be imposed without requiring analysis of supplementary samples.

Animals↗
PubMed1995

[The use of ELISA and ultramicro ELISA in identifying a Coxsackie A9 strain isolated during the epidemic of neuropathy in Cuba].

During 1993 an outbreak of epidemic neuropathy characterized by 2 major clinical forms: the optic neuropathy and peripheral neuropathy, occurred in our country. Although the cause of the disease was considered to be multifactorial (mainly due to neurotoxic agents and to nutritional deficiency) an Enterovirus was isolated from the cerebrospinal fluid of a patient (strain 47/93, IPK) which was subsequently classified as a Coxsackie A9 virus (Cox A9) by the neutralization test, and using pools of Lim Benyesh Melnick (LBM) serum. Results obtained during the application of ELISA and ultramicro ELISA techniques for the identification of this agent from the supernatant of a culture of infected cells are reported, as well as the detection of antibodies to the strain found within populations with high and low incidence rates of the disease.

Antibodies, Viral↗
PubMed2006

A sensitive and reliable quantification method for mouse interleukin-12 p70 based on fluorometric sandwich ELISA (FS-ELISA).

Interleukin-12 (IL-12) p70 is an important cytokine secreted by antigen-presenting cells in response to antigenic stimulation; it is a heterodimer of p35 and p40 subunits. Here, we report a new, highly sensitive, and reliable method that employs fluorometric sandwich ELISA for quantification of the mouse IL-12 (moIL-12) p70 protein. Our method could quantify moIL-12 p70 in the range of approximately 0.5 to 500 pg/ml. In the assay, no signals were produced by the moIL-12 p40 monomer, homodimer [(p40)2], or mouse IL-23 even up to a concentration of 500 pg/ml; this ensures that our assay has a high specificity for moIL-12 p70. To demonstrate that our method can determine natural moIL-12 in real physiological/pathological samples, we monitored the moIL-12 p70 secretion from peritoneal exudative cells after lipopolysaccharide (LPS) stimulation. IL-12 p70 secretion as early as 3h after LPS stimulation was reliably detected due to the high sensitivity of the method.

Animals↗
PubMed2005

Analysis of Borrelia burgdorferi IgG antibodies with a combination of IgG ELISA and VlsE C6 peptide ELISA.

Enzyme-linked immunosorbent assays (ELISAs) were used to detect antibodies to the C6 peptide of the Borrelia burgdorferi VlsE protein and a selection of B. burgdorferi IgG antigens, separately and as a combination, in 355 serum specimens from blood donors and patients. Western immunoblotting was used as the reference method. The sensitivity of the combined analysis of IgG antigen and C6 peptide analysis was markedly superior to those of the separate analyses. When the C6 peptide and IgG results were concordant, the customary confirmatory Western immunoblotting assay could be omitted, thus reducing the time and cost of analysis.

Antibodies, Bacterial↗
PubMed1990

Studies on the development of an ELISA kit for microbiological monitoring. 2. Improvement of the prototype ELISA kit with special references to mouse hepatitis virus antigen.

Improvement of the mouse hepatitis virus (MHV) antigen in a prototype ELISA kit was performed. Equivalent divalent antigens of MHV Nu-67 and S strains with a protein concentration of 10 micrograms/ml showed the best sensitivity and specificity for the detection of MHV and sialodacryoadenitis/Parker's rat coronavirus antibodies in mice and rats, respectively. An increase in the reliability of macroscopic evaluation of both antibody tests is expected by using the newly manufactured kit with the improved antigen.

Animals↗
PubMed2000

Assessment of gastric carcinoma risk associated with Helicobacter pylori may vary depending on the antigen used: CagA specific enzyme-linked immunoadsorbent assay (ELISA) versus commercially available H. pylori ELISAs.

BACKGROUND: Previous epidemiologic studies produced inconsistent results when examining the relation between Helicobacter pylori infection and the risk of gastric carcinoma by measuring various anti-H. pylori antibodies. This study investigated the increased risk of cancer by examining different antibodies, including the specific anti-CagA antibody and antibodies from two commercially available kits. METHODS: An ELISA for the detection of serum anti-CagA was established using a recombinant CagA protein that the authors previously reported. Serum anti-CagA titer was determined for 80 patients with gastric carcinoma and 80 gender- and age-matched controls. Two anti-H. pylori antibodies from the commercially available kits HEL-p (Amrad, Kew Vic, Australia) and HM-CAP (Enteric Product Inc., Westbury, NY) were also evaluated. RESULTS: Anti-CagA seropositivity differed significantly between gastric carcinoma patients and controls (92.5% vs. 55.0%; P = 0. 0001), showing an odds ratio of 10.4 (95% confidence interval [CI]: 4.23-29.74). The difference was less prominent for the seropositivity of HEL-p (77.5% vs. 58.8%; P = 0.0139; odds ratio: 2. 38; 95% CI: 1.20-4.82) and insignificant for that of HM-CAP (65.0% vs. 57.5%; P = 0.4325; odds ratio: 1.30; 95% CI: 0.68-2.49). CONCLUSIONS: The current study revealed that the antibody assay system used could be one important factor in the assessment of gastric carcinoma risk for patients with H. pylori.

Adult↗
PubMed1978

A new technique of heterogenous enzyme-linked immunosorbent assay, stick-ELISA. II. Application of stick-ELISA to antigens of various infective agents.

Stick-enzyme-linked immunosorbent assay (Stick-ELISA) was applied to antigens of a variety of infective agents. An acceptable reproducibility in a series was reached, the variation coefficient being twelve percent. Preliminary results revealed that ready coated sticks could be stored at 4 degrees C for at least two to three weeks.

Antigens↗
PubMedDate not supplied

Determination of aflatoxin concentrations in peanut butter by enzyme-linked immunosorbent assay (ELISA): study of three commercial ELISA kits.

Sixteen United Kingdom analytical laboratories participated in an evaluation of 3 commercially available enzyme-linked immunosorbent assay (ELISA) kits for analysis of aflatoxin in peanut butter. Each laboratory was sent 3 sets of 10 randomly numbered samples of peanut butter. Each set consisted of 5 pairs of undisclosed duplicates. Four of the sets of duplicates were naturally contaminated butters with "target" aflatoxin values (estimated by liquid chromatography) between 8 and 81 micrograms/kg. The fifth pair was a blank peanut butter containing approximately 3 micrograms/kg of total aflatoxins. A statistical treatment of the results of the study is presented, together with discussion of the relative merits of the different kits.

Aflatoxins↗
PubMed1988

Detection and quantification of anti-HBs antibodies by the Czechoslovak ELISA SEVAC Kit (Sevatest ELISA anti-HBs).

We invented and verified the possibilities of clinical use of the ELISA Kit for quantification of human serum anti-HBs antibodies. The kit does not employ any labelled antigen but is based on the principle of neutralization. The kit was tested on the panel of sera of the normal population, blood donors, workers at risk workplaces and persons vaccinated with Heptavax-B vaccine. The sensitivity of the kit is 80 mIU/ml, its capture is only slightly worse (in the convalescents) than the capture of the AUSAB (Abbott, USA). Statistical values and frequency distribution at various levels of anti-HBs antibodies in the followed-up group, have been indicated. The kit can be used very well for the follow-up of patients afflicted with virus hepatitis B, for the observation of workers at risk workplaces, for selection of persons eligible for vaccination and for checking the vaccination. It is suitable for the selection of sera for the preparation of specific globulin against hepatitis B and for standardization of this preparation. It is less suited for the compilation of epidemiological surveys where it is important to detect even as low levels of antibodies as 10-80 mIU/ml.

Blood Donors↗
PubMed1993

Detection of the Haemophilus somnus antibodies in the bulls' reproductive tract fluids using the ELISA. I. Elaboration of the ELISA for the detection of the specific antibodies in the IgG, IgM and IgA classes.

The conditions of the ELISA for detecting the Haemophilus somnus antibodies in IgG, IgM and IgA classes were elaborated. The test was adapted for examining the seminal plasma, preputial washings and blood serum samples of bulls. In order to obtain the more precise evaluation of results, the tests were made for two different dilutions of the examined material. The arising out of this method difficulty in the evaluation of the intensity of reaction was solved by introducing the specific, semi-quantitative method of classification, using the eleven-degree scale. The mean arithmetic classificational values calculated from absorbance readings were called "absorbance index". The introduced parameter proved to be especially useful while comparing the reaction of antibodies in reproductive tract fluids samples.

Animals↗
PubMed1995

Quantitative measurement of HBeAg in chronic hepatitis B: a comparison between a radioimmunoassay, a fluorescence ELISA and a chemiluminescence ELISA.

The presence of the hepatitis B e antigen (HBeAg) in peripheral blood of chronic hepatitis B patients is a widely accepted marker of active replication of the hepatitis B virus. HBeAg determination during interferon therapy is a useful guide for the therapeutic regimen. The aim of the study was to compare the suitability of an HBeAg radioimmunoassay (RIA, Abbott Laboratories, North Chicago, IL, USA), the IMx-HBeAg assay (IMx, Abbott Laboratories) and the HBeAg/anti-HBe Amerlite assay (Amerlite, Johnson & Johnson Clinical Diagnostics, Cardiff, UK) for semiquantitative monitoring of HBeAg during therapy. HBeAg levels in serum samples obtained before and during interferon therapy were measured using an in-house standard calibrated against the Paul Ehrlich Institute HBeAg reference preparation (PEI standard). When serial dilutions of pretreatment serum samples were assayed by the three methods, radioimmunoassay was found to be highly sensitive although it had a very limited working range (0.5 to 12 PEI U/ml). A broader linear working range was observed for Amerlite (0.5 to 50 PEI U/ml) and the IMx assay (0.5 to 100 PEI U/ml). The intra-assay and interassay variations did not differ significantly. Since the IMx assay was less susceptible to sample variation and had a broad working range, semiquantitative measurement of HBeAg in one diluted and one undiluted sample by this assay may justifiably be introduced as routine procedure. Routine semiquantitative HBeAg measurement may improve individual dose adjustments and thus the success of interferon therapy.

Chronic Disease↗