Multicenter Study on Incubation Conditions for Environmental Monitoring and Aseptic Process Simulation.

Unlabelled: Environmental monitoring and aseptic process simulations represent an integral part of the microbiological quality control system of sterile pharmaceutical products manufacturing operations. However, guidance documents and manufacturers practices differ regarding recommendations for incubation time and incubation temperature, and, consequently, the environmental monitoring and aseptic process simulation incubation strategy should be supported by validation data. To avoid any bias coming from in vitro studies or from single-site manufacturing in situ studies, we performed a collaborative study at four manufacturing sites with four samples at each location.

Yeast identification in the clinical microbiology laboratory: phenotypical methods.

Emerging yeast pathogens are favoured by increasing numbers of immunocompromised patients and by certain current medical practices. These yeasts differ in their antifungal drug susceptibilities, and rapid species identification is imperative. A large variety of methods have been developed with the aim of facilitating rapid, accurate yeast identification.

Evaluation of six commercial systems for identification of medically important yeasts.

Six commercially available systems for the identification of yeasts were evaluated using 133 clinical isolates and four reference strains that had been previously identified by conventional methods and 19 recent clinical isolates that had been identified by the ID32C system (bioMérieux, France). The total identification rates (TIR) established for the total number of strains tested and the database identification rates (DBIR) established for the strains included in the respective manufacturer databases were both determined. After incubation for 4 h, the TIR and DBIR were 78% and 84%, respectively, for the RapID Yeast Plus system (Innovative Diagnostic Systems, USA).

Survey of primary drug resistance of Mycobacterium tuberculosis in Casablanca, Morocco.

Setting: In 1990, a 6-month short-course regimen (2 SHRZ/4 RH) was introduced for the treatment of tuberculosis in Morocco.

Objective: To assess the efficacy of the national tuberculosis control programme, a prospective study of primary drug resistance was conducted from April 1992 to July 1994 in Casablanca.

Design: A total of 402 strains isolated from 402 patients living in Casablanca with no previous history of tuberculosis was included in the study.

[Chlamydia infection and female low fertility in Morocco].

Chlamydia trachomatis infection is recognised as the most common asymptomatic sexually transmitted disease, and this may lead to severe complication including infertility. The purpose of this study was to evaluate the part that this pathology takes in the female hypofertility, using serologic, cell culture, and histopathologic tests. Some of the women had undergone biopsies during coelioscopic exam, the others during salpingectomy.

[Chlamydial infections and male infertility in Morocco].

Chlamydia trachomatis infection of the lower genital is recognised as the most common sexually transmitted disease, is role in male infertility is controversial, the objective of this study was to evaluate the part that this pathogen agent takes in male infertility among maroccan population, to compare serological tests, sperm abnormalities, antisperm-antibodies and DNA research in semen. Microimmunofluorescence (MIF) was done for 139 patients, 124 were checked for sperm abnormalities, 87 for antisperm-antibodies and 92 for DNA research in sperm. The results showed that MIF is positive in 24,5%, 11% of the subjects in antisperm antibodies, 8% of them simultaneously in anti-Chlamydia and antisperm antibodies and 5 of them had sperm abnormalities.

[Isolation of Chlamydia trachomatis in trachomatous Moroccan patients].

Purpose: Isolation of Chlamydia trachomatis from ocular specimens of subjects living in trachomatous area in south Morocco.

Methods: One hundred and twenty ocular specimen of 60 subjects living in two provinces of a trachoma-endemic area (Errachidia and Ouarzazate) were tested by cells culture. The age range was 2 months to 85 years and the sex ratio was 1.

Production of a new monoclonal antibody specific to human des-gamma-carboxyprothrombin in the presence of calcium ions. Application to the development of a sensitive ELISA-test.

In order to explore pathologies possibly associated with vitamin K deficiency, several monoclonal antibodies (mAbs) were produced against human Desgamma-Carboxy-Prothrombin (DCP). One of these mAbs, designated C4B6, detected DCP forms in the presence of Calcium ions, confirmed by comparison with the patterns of two electrophoretic techniques: Affino-Immuno-Electrophoresis (CAIE) and Polyacrylamide Gel Electrophoresis followed by Electro-blotting (PAGE-Blot). An Enzyme-Linked-Imunosorbent Assay (ELISA) using mAb C4B6 has been developed, optimized and standardized.

[Seroprevalence of Chlamydia trachomatis infection in STD consultants in Morocco].

We have conducted a seroepidemiological survey of Chlamydia trachomatis infection among 400 STD consultants in comparison with 400 blood donors. The study was performed by using the indirect microimmunofluorescence technique with Chlamydia trachomatis, Chlamydia psittaci and Chlamydia pneumoniae as antigens. The overall seroprevalences were 60% and 46% for STD consultants and blood donors respectively.

[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.

Evaluation of five commercial antifungal susceptibility testing systems.

Five commercial antifungal susceptibility testing systems were studied for repeatability and reproducibility as well as concordance of results with the MICs for ten reference strains belonging to six different species. Repeatability was determined by testing each strain in triplicate on the same day, and reproducibility by repeating this triple determination on three different days. On the basis of 630 yeast-antifungal agent results for Mycototal and Mycostandard, 540 for Candifast, and 450 for ATB Fungus and Diff Test, repeatability was consistently equal to or greater than 95%.

Purification and immunological studies of the cross-reaction between the 65-kilodalton gonococcal parietal lectin and an antigen common to a wide range of bacteria.

The 65-kDa gonococcal parietal lectin (GPL) has been purified and found to have a lectinlike activity exhibiting both structural and immunological similarities to the common antigen family. Ultrastructural localization of GPL was done by using anti-GPL monoclonal antibodies: GPL is a major component of the outer membrane and is also present in blebs formed by gonococci.

Three electrophoretic techniques comparison for des-gamma-carboxyprothrombin detection.

Des-gamma-carboxyprothrombin (DCP) is a marker that appears in the blood when modifications of vitamin K-dependent proteins carboxylation cycle occur. About 280 human plasma samples of diverse origins were tested by three different electrophoretic techniques for the evaluation of DCP: rocket immunoelectrophoresis (RIE) before and after barium carbonate adsorption, crossed affinoimmunoelectrophoresis (CAIE) and polyacrylamide gel electrophoresis in presence of calcium lactate followed by immunoblotting (PAGE-blot). A good correlation was found between CAIE and PAGE-blot in the CAIE detection limit, but not between RIE and the two other techniques.

Protein A insensitive ELISA detection of staphylococcal enterotoxin B.

A sandwich enzyme-linked immunosorbent assay to detect staphylococcal enterotoxin B was developed using rat monoclonal antibodies as capture antibodies and as a biotinylated conjugate. This test was sensitive, less than 1 ng/ml of enterotoxin B was detected and interference by protein A was prevented by the use of rat monoclonal antibodies of the IgG2a isotype which were insensitive to protein A even at concentrations greater than 1000 ng/ml.

Des-gamma-carboxyprothrombin detection by immunoblotting after polyacrylamide gel affinoelectrophoresis in human plasmas.

In the absence of vitamin K or in the presence of the vitamin K antagonists, abnormal nonfunctional forms of prothrombin circulate in the blood. A reliable and reproducible technique, derived from traditional crossed affinoimmunoelectrophoresis in presence of calcium lactate, was developed and optimized. The technique is based on nondenaturing polyacrylamide gel affinoelectrophoresis, with calcium lactate, of plasma samples, followed by immunoblotting with rabbit anti-human prothrombin serum and detection with an anti-rabbit immunoglobulin peroxidase conjugate.

Rapid identification of Candida albicans by dot-enzyme immunoassay.

A highly sensitive and specific dot-enzyme immunoassay for the rapid identification of Candida albicans was developed using a murine monoclonal antibody (Mab), which adsorbed to cell surface-exposed determinants. This Mab reacted with 28 of the 28 C. albicans strains tested including the serotypes A and B and 2 C.

Comparison of sandwich-ELISA and GM1-ELISA for the detection of Escherichia coli thermolabile enterotoxin.

Two different microtiter plate ELISA tests were devised for the detection of Escherichia coli thermolabile toxin (LTh) either free or extracted from isolated colonies. Both tests used as detection systems purified anti-LTh rabbit immunoglobulins conjugated to biotin, streptavidin peroxidase and TMB. The tests differed by their capture phase which was the GM1-ganglioside for GM1-ELISA and purified anti-LTh rabbit immunoglobulins for sandwich ELISA.

Characterization of Mycobacterium species by sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

A rapid method has been standardized for extraction and SDS-PAGE analyses of mycobacterial cellular protein. Complex patterns were reproducibly obtained of the 31 reference strains or clinical isolates studied, belonging to 11 species. These patterns were species specific; thus this method could be an easy tool for mycobacterial strain characterization.

[Comparative sensitivity of yeasts to ketoconazole, itraconazole and fluconazole using a liquid medium standardized micromethod].

The minimal inhibitory concentrations of 200 yeast strains (48 reference strains and 152 recently isolated from pathological products) were evaluated with a new standardized micromethod using a liquid medium comparatively for ketoconazole, itraconazole and fluconazole. The ready-to-use microtitration plates contained the antifungal agents in concentrations ranging from 100 to 0.10 mg/l.

[Sensitivity of Aspergillus to ketoconazole and itraconazole using a standardized micromethod].

The standardized micromethod Mycototal is based on ready-to-use microtitration plates and the same culture medium for sensitivity testing of all antifungal agents. Since the method was very reproducible for yeasts it was applied for the determination of minimal inhibitory concentrations of Aspergillus to itraconazole and ketoconazole. Again the reproducibility for itraconazole was excellent since for 19 strains tested two times 18 showed no more than two dilutions different results and this was also observed for 6 strains over 7 tested 4 or 5 times.

[Fast identification of Streptococcus pyogenes serotype M.12 in throat swabs by an improvised coagglutination technique].

A rapid procedure for identification of Streptococcus pyogenes serotype M. 12 directly in throat swabs, is reported and compared with standard culture method on blood agar plates and typing of group A Streptococci isolated, with double gel immuno-diffusion. This procedure consist of chlorhydric acid extraction of swabs and testing of the extract towards specific M.