Thin layer chromatography characterization of ELISA specific glycolipids antigens immunomagnetically purified from environmental mycobacteria.

Thin layer chromatography (TLC) could easily and rapidly evidentiate the qualitative differences between glycolipids (GLs). Different immunomagnetically purified mycobacterial GLs have been compared using TLC, in order to choose the most appropriate antigens to be utilized in ELISA. The GLs were purified from environmental mycobacteria (EM) (M.

Use of recombinant purified protein derivative (PPD) antigens as specific skin test for tuberculosis.

Background & Objectives: Purified protein derivative (PPD) is currently the only available skin test reagent used worldwide for the diagnosis of tuberculosis (TB). The aim of this study was to develop a Mycobacterium tuberculosis specific skin test reagent, without false positive results due to Bacillus Calmette-Guerin (BCG) vaccination using recombinant antigens.

Methods: Proteins in PPD IC-65 were analyzed by tandem mass spectrometry and compared to proteins in M.

Prospective Comparison of Two Brands of Tuberculin Skin Tests and Quantiferon-TB Gold in-tube Assay Performances for Tuberculosis Infection in Hospitalized Children.

Objective: The study compared two brands of tuberculin skin tests (TST): PPD RT 23, (SSI, Denmark) and PPD IC 65, (Cantacuzino Institute, Romania), 2 TU/ 0.1 ml each, with an interferon gamma release assay [IGRA], Quantiferon-TB Gold (QFT).

Material And Methods: QFT was performed on whole blood samples, before TSTs, on 60 children with tuberculosis (TB), BCG vaccinated, admitted in a paediatric pneumophtisiology hospital.

Serodiagnosis of environmental mycobacterial infections.

To demonstrate the usefulness of enzyme-linked immunosorbent assay for serodiagnosis of mycobacterioses due to environmental mycobacteria we utilized a panel of glycolipid antigens selective for Mycobacterium avium-intracellulare, Mycobacterium kansasii, Mycobacterium xenopi, Mycobacterium scrofulaceum and Mycobacterium gordonae. The levels of circulating antibodies were determined against the environmental mycobacteria, and Mycobacterium tuberculosis in human immunodeficiency virus-negative and -positive patient sera. The method used immunomagnetic separation of the antigens, with covalent immobilization of antibodies to superparamagnetic amine and carboxyl terminated particles in solutions of the specific antigens.

Comparative study of RT23 and IC-65 tuberculins tested on children with tuberculosis.

This study, conducted in 2009, proposed to evaluate and compare the biological potency of two different tuberculins, RT23 (Statens Serum Institute, Copenhagen) and IC-65 (Cantacuzino Institute, Bucharest) when administered to 89 children with confirmed tuberculosis, admitted to Paediatric Department of Pneumophtysiology Institute, Bucharest. Mean age of subjects was 10.4 years [SD (standard deviation) = 5.

Comparison of tuberculin skin test with a whole-blood interferon gamma assay and ELISA, in HIV positive children and adolescents with TB.

We compared the usefulness of three methods designed to diagnose latent tuberculosis [TB]: interferon-gamma release assay [IGRA], such as QuantiFERON-TB Gold [QFT-G], Enzyme-linked immunosorbent assay [ELISA] serologic assay and tuberculin skin test [TST] for diagnosis of TB in human immunodeficiency virus [HIV]-1 infected children and adolescents, with microbiologically and/or histopathologically confirmed TB co-infection. The serum samples were obtained from 36 patients who were examined and tested by the three methods. The sensitivity was 38.

Establishing of a minimal set of tests (minimal standards) to identify species included in the genus Mycobacterium.

Mycobacterium genus includes over 100 species and subspecies; new species are discovered every year. Minimal standard criteria are represented by the resistance to acid-alcohol (e.g.

Rapid immunochromatographic serum assay of nontuberculous mycobacterial infections.

A rapid immunochromatographic serologic assay (Dot assay) is proposed to be applied on patients infected with nontuberculous mycobacteria (NTM). This assay could evidentiate the infecting species and allow the beginning of the treatment. The test is based on the principle of immunoblotting chromatography, a rapid membrane-based assay, capable of diagnosing NTM infections in serum, in less than 1 hour, with no need of special equipment or skilled staff.

Rapid identification of Mycobacterium species by lectin agglutination.

The purpose of the present study is to explore the possibility that plant lectins can be used for the development of rapid and inexpensive technique for differentiation of mycobacterial species. The method is based on interaction between mycobacteria and lectins as visualized by agglutination in a microtiter plate. We employed 18 mycobacterium species and determined the minimal lectin concentration (MLC) of 23 different lectins.

Rapid dot sputum and serum assay in pulmonary tuberculosis.

A rapid direct sputum (Sp.) and/or antibody assay, based on immunoblotting and enzyme immunoassay is described. The test can detect mycobacterial antigens or antibodies in clinical specimens from pulmonary tuberculosis (TB) patients.

Serodiagnosis of extrapulmonary tuberculosis by enzyme-linked immunosorbent assay (ELISA).

Antibodies against Mycobacterium tuberculosis antigenic glycolipids were determined by enzyme-linked immunosorbent assay (ELISA). The 720 sera were collected from adult patients under investigation, suspected with extrapulmonary tuberculosis. The test performance was estimated according to definitive diagnosis in terms of specificity, sensitivity, positive predictive value and negative predictive value.

Hemagglutinin of unusual specificity from Helcococcus kunzii.

Helcococcus kunzii is a gram-positive, catalase-negative opportunist. The organism has been isolated from the lower extremities and breast masses of several patients. A clinical isolate of Helcococcus kunzii was shown to possess a hemagglutinin-lectin with a specificity for N-acetylglucosamine and lactose, two structurally unrelated carbohydrates.

Application of ELISA with antigenic glycolipids in early diagnosis of pulmonary tuberculosis.

Antibodies against M. tuberculosis antigenic glycolipids were determined by enzyme-linked immunosorbent assay (ELISA) in 80 sera from patients suspected of pulmonary tuberculosis, with suggestive clinical signs and radiological abnormalities, but smears negative. The test was also performed on 68 control sera from patients with bacteriologically confirmed pulmonary tuberculosis, in different stages of disease.

Serodiagnosis of tuberculous meningitis by enzyme-linked immunosorbent assay (ELISA).

IgG antibodies against glycolipids and proteins isolated from M. tuberculosis and BCG suspension were determined by ELISA in sera, in CSFs and in serum and CSF paired samples, from patients with tuberculous meningitis and from healthy control subjects. With specificities between 90 and 94% for the antigens used, we obtained senitivities of 75% for Pr-ELISA, 60% for G1-ELISA and 35% for BCG-ELISA.

Clinical usefulness of rapid serodiagnosis in pulmonary tuberculosis by ELISA with glycolipidic antigens extracted from Mycobacterium tuberculosis and with whole BCG suspension.

Antibodies against Mycobacterium tuberculosis glycolipids and whole BCG suspension were determined by ELISA on 58 sera from hospitalized patients with presumptive pulmonary tuberculosis and on 127 sera from control subjects. The experimental results demonstrated that the glycolipids are more adequate to be used as antigens than whole BCG suspension, as high sensitivity of the test was obtained. By using only one antigen, ELISA becomes more efficient in rapid diagnosis.

Humoral immune response in human pulmonary tuberculosis: antibodies against Mycobacterium tuberculosis polysaccharide, protein and glycolipid antigens.

Sera from 67 tuberculosis patients and 30 healthy subjects have been analyzed for the presence of specific antibodies against polysaccharide, protein and glycolipid antigens from H37Rv strain of Mycobacterium tuberculosis. The results proved the advantages of using the glycolipid antigens in diagnosing tuberculosis as well as the relationship between the antibodies level and the extension of the pulmonary tuberculous lesions.

Interleukin 2 enhances the efficiency of immunotoxins in the treatment of mice with ascitic tumors.

Two multivalent immunotoxins (ITs) with cytotoxic potential against Thy 1.2-expressing tumor cells were used in association with mouse interleukin 2 (IL2) for treatment of mice bearing ascitic EL4 lymphomas. The combined treatment, ITs + IL2, induced an enhanced antitumor effect revealed by a significant prolongation of the survival time of mice as compared to the simple treatment with ITs or IL2 alone.

Specific antibodies and mycobacterial antigens in patient sera with pulmonary tuberculous and nontuberculous diseases. Note II.

"Two-assay" tests (TAT), immunoenzymatic determination of both specific antibodies and mycobacterial antigens in sera of tuberculous and non-tuberculous subjects, was undertaken in our territorial conditions, where BCG vaccination is systematically applied and the prevalence of tuberculous infection is relatively high. The sensitivity of the method, calculated on 42 patients with active pulmonary tuberculosis and on 39 patients with post-tuberculosis syndromes is high, i.e.

Specific antibodies and mycobacterial antigens in pulmonary tuberculosis patients sera. Note I.

206 sera collected from different groups of subjects were analyzed by immunoenzymatic methods regarding the content of Mycobacterium tuberculosis specific antibodies and mycobacterial antigens. The results underlined that two assays offered improved serologic diagnosis of tuberculosis over a single antibody test. BCG vaccination interferes with serologic tests for tuberculosis when polyspecific antibodies or mixture of common and specific antigens are used as immunologic reagents.

Treatment of murine EL4 leukemia in ascitic form with anti-Thy 1.2 specific immunotoxins.

C57BL/6 mice with EL4 leukemia cells in ascitic form were intraperitoneally treated with ricin A chain-multivalent antibody immunotoxins. The immunotoxins containing rabbit IgG anti-Thy 1.2 antibodies complemented by protein A of Staphylococcus aureus were able to interact specifically with the target cells and to induce an antitumor effect as revealed by an increase in survival time of the mice.