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J Orfila

Publications and source records attributed to J Orfila.

At least 37 records · Page 2Linked to original sources
PubMed1999

[Prevalence of anti-Chlamydia pneumoniae antibodies in preadolescent children in Congo].

Chlamydia pneumoniae (CPn) is recognised as a common cause of both upper and lower respiratory tract infections. Seroepidemiological studies seem to indicate a world-wide distribution of this organism. In order to evaluate the prevalence of antibodies to CPn in a healthy pediatric population, we measured anti-C. pneumoniae antibodies in a group of 253 infants without respiratory tract infections, aged from 1 to 12 years. Sera were obtained from children seen at immunization clinics and schools in Lubumbashi (Congo). Antibodies to CPn were evaluated using micro-immunofluorescence assay. IgG antibody to CPn in a titre 16 was considered as positive. The antibody prevalence was found to be 25.7%. This prevalence was 6.2% in children aged from 1 to 6 years, and 37.8% in children aged from 7 to 12 years. It was less than 10% under five years, increasing to 50% at 12 years. The progressive increasing of seropositivity related to age suggests that reinfections may be frequent. This study shows an important spread of this bacteria in the preadolescent population of an African country.

Antibodies, Bacterial↗
PubMed1997

Capture-ELISA: a new assay for the detection of immunoglobulin M isotype antibodies using Chlamydia trachomatis antigen.

We present here a new method of IgM antibody-capture enzyme-linked immunosorbent assay (IgM-Capture-ELISA) for the diagnosis of recently acquired infections with Chlamydia trachomatis. For this analysis, plates were coated with goat IgG anti-human Fc mu. The capture of serum IgM antibodies was revealed indirectly by the sequential addition of biotinylated chlamydial proteins and peroxidase-conjugated streptavidin. In chlamydial extracts, cysteine-rich proteins are preferential antigenic targets for the humoral response. 3-(N-maleimidopropionyl)-biocytin (MPB), which binds biotinylated moieties to sulfhydryl groups, was used for the labeling procedure. The preservation of the antigenic specificity of labeled proteins was controlled by a blotting of these proteins, which were, respectively, probed either with specific IgM antibodies or with streptavidin. This analysis revealed that, after labeling, recognized epitopes are more particularly present on the major outer membrane protein (MOMP) of Chlamydia trachomatis. The validation of IgM-Capture-ELISA was assessed by using 170 selected sera from patients suspected of being infected by Chlamydia. Results were respectively compared to conventional indirect immunofluorescence assays (MIF-IgM assays) and to Western blotting. Sixteen sera were found to possess IgM antibodies against Chlamydia trachomatis with IgM-Capture-ELISA. Among these 16 sera, 14 and 15 were, respectively, positive with MIF-IgM assays and in Western blotting. Data obtained with IgM-Capture-ELISA reveal the absence of false-positive results in sera containing rheumatoid factor, which has been shown to interfere in the two other methods. IgM-Capture-ELISA value was then confirmed using sera from patients consulting for genital or pulmonary diseases, from patients with confirmed chlamydial infections, and from patients with other pathologies. IgM-Capture-ELISA appears as an alternative simple semi-quantitative assay for the detection of early chlamydial infection.

Animals↗
PubMed1997

Genotyping of Portuguese Chlamydia trachomatis urogenital isolates.

OBJECTIVE: To determine the prevalence of the different Chlamydia trachomatis genotypes in Portuguese patients. METHODS: Urogenital isolates (n = 240) derived from attenders of various clinics in the Lisbon area were differentiated into genovars by genotyping with restriction fragment length polymorphism (RFLP) analysis of the PCR amplified omp1 gene. RESULTS: Genotype E was the most common for both men (47.9%) and women (43.8%). Genotypes D and F were the second most prevalent for men (11.3%) and genotype H was the second most prevalent for women (19.5%). Genotypes F, G, D, in women and H, G, I, in men, were found in a lower percentage of cases. Genotypes B, Ba, J, K, L1 and L2 were very rarely detected. CONCLUSIONS: With one exception, the overall distribution of Chlamydia trachomatis genotypes in our study is similar to what has been observed in other western countries. The only exception is the unusual prevalence of genotype H among women. The clinical manifestations associated with this and other genotypes were similar.

Adult↗
PubMed1997

[Research of type 1, 2 and 3 hepatitis C virus specific antibodies in relation to the viral genotype in chronically infected patients].

The determination of the viral genotype in chronically HCV infected patients is used as an epidemiologic tool, a predictive marker of the evolution of the disease and especially of the response to the interferon alfa therapy. Its determination needs an amplification of the selected genomic region and remains expensive. Thus an indirect determination of the genotype has been proposed using the characterization of type-specific antibodies. In 101 chronically HCV infected patients, a serologic method specific for HCV type 1, 2 and 3 has been evaluated in relation to the genotyping one. The test was interpretable in 64 sera and in agreement with the genotyping method in 92% of the samples. The reactivity of the test was lower in hemodialysis and IVDUs patients (p < 0.02). The mixed results obtained by the serologic typing method were not in agreement with the results of the genotyping coinfections. We could not differentiate the possible past infections from the cross-reactivities of the test. The serotyping methods was simple and rapid, allowing generally a deduction of the viral genotype. It could be an attractive approach if the reactivity was improved and the subtyping was possible.

Adult↗
PubMed1997

[Use of an experimental Chlamydia trachomatis salpingitis model for evaluating the effectiveness of antibiotics and anti-inflammatory agents on fertility].

OBJECTIVES: An experimental model of Chlamydia trachomatis salpingitis was developed in mice to study the effect of combination antibiotic and antiinflammatory therapy on restoring fertility after tubal infection. METHODS: The mice were infected by injection via the ovarian bursa with a suspension containing a human strain of Chlamydia trachomatis sevar F. After treatment with antibiotics alone or in combination with antiinflammatory agents, fertility was studied at 4 months. RESULTS: With doxycycline and ofloxacine, there was not a significant difference in fertility between animals treated with antibiotics alone and those treated with antibiotics and antiinflammatory agents. However, a piroxicam and azythromycin combination improved fertility with a longer delay than with azythromycin alone. The most important factor appeared to be the antibiotic treatment. CONCLUSION: Associating an antiinflammatory agent was not shown to have a beneficial effect in reducing infertility after Chlamydia trachomatis salpingitis.

Animals↗
PubMed1996

Limits of the microimmunofluorescence test and advantages of immunoblotting in the diagnosis of chlamydiosis.

The Western blot (immunoblot) patterns of 56 serum specimens, all examined previously by the microimmunofluorescence (MIF) test for species-specific Chlamydia antibodies, were analyzed. Predominant specific-antibody activity was directed to the 170-, 155-, 145-, 120-, 115-, 100-, 57-, and 38-kDa proteins of Chlamydia trachomatis and to the 175-, 130-, 110-, 98-, and 30-kDa proteins of Chlamydia pneumoniae. All of these antigens appeared to be species specific. The reactivity with 90-, 80-, 75-, 62- or 60-, and 55-kDa proteins and the major outer membrane protein appeared to be genus specific. Fourteen serum samples which had identical titers of immunoglobulin G as determined by the MIF test were investigated by Western blotting. We found that nine serum samples had anti-C. trachomatis protein profiles and two had anti-C. pneumoniae protein profiles. The double seropositivity observed by MIF corresponded with cross-reactivity to genus-specific antibodies. As for the three remaining serum specimens, we observed identical protein profiles for C. trachomatis and C. pneumoniae, confirming the double seropositivity experienced with the MIF test. Western blotting can differentiate between specific reactions and interfering noise from other, partly cross-reacting chlamydial species.

Antibodies, Bacterial↗
PubMed1995

Denaturing gradient gel electrophoresis analysis for the detection of point mutations in the Chlamydia trachomatis major outer-membrane protein gene.

Fifty clinical strains of Chlamydia trachomatis were studied by denaturing gradient gel electrophoresis (DGGE) of bacterial DNA amplified by the polymerase chain reaction (PCR). The strains belonged to the three most commonly encountered serovars in developed countries--D, E and F. Six reference strains, including the serovar Da strain, were also studied. The DNA sequences explored encompassed the four variable domains (VDs) of omp1, the gene encoding the major outer-membrane protein (MOMP). The corresponding regions in the MOMP contain the species-, subspecies- and serovar-specific epitopes. The four distinct serovars were clearly differentiated by specific migration pattern. No sequence variations were observed among strains of serovar F. However, variant strains within serovars D and E were found, which exhibited migration patterns different from those of the reference strains and these were sequenced directly. According to the observed sequence variations, serovar D strains could be divided into three stable representative groups (D, D1 and D2). Two variants were identified among serovar E strains. No biological differences were observed for the variant strains in terms of growth properties, ecology or pathogenicity. All the nucleotide substitutions detected in the VDs were non-synonymous at the protein level and, for the serovar D strains, could account for differences identified by specific monoclonal antibodies. These substitutions could be involved in antigenic drift, driven by the immune pressure of the host, leading to the emergence of serovariants. The data may explain, in part, chlamydial infection recurrences and could have critical implications for developing rational vaccine strategies.

Algorithms↗
PubMed1995

Heterogeneity within the first constant segment of the major outer membrane protein gene in Chlamydia trachomatis serovar D/Da distinguishes 2 lineages.

Restriction fragment length polymorphism of the major outer membrane protein gene (ompl) was studied in 73 epidemiologically unrelated Chlamydia trachomatis serovar D (n = 64) and Da (n = 9) strains isolated between 1983 and 1991 in various european countries from patients suffering from sexually transmitted diseases, as well as 3 reference strains D/IC-Cal-8 (USA), D/UW3 (UK) and Da/TW-448 (Taiwan). Among these strains, 5 different genotypic groups were distinguished: 3 for the serovar D strains and 2 for the serovar Da strains. Comparisons of the complete ompl nucleotide sequence of 1 member of each group revealed 2 distinct lineages, independently of the D/Da classification, presumably as the result of an intragenic DNA recombination event within the first constant segment. The observed ompl genetic diversity of serovar D/Da strains in regions involved in T- and B-cell responses might add another level of complexity to the design of a subunit vaccine.

Amino Acid Sequence↗
PubMed1994

Sexually transmitted agents and cervical neoplasia in Colombia and Spain.

Case-control studies of cervical intra-epithelial neoplasia grade III (CIN III) and of invasive cervical cancer were carried out in Spain and Colombia to assess the relationship between cervical cancer and 6 common sexually transmitted agents (STAs). The CIN-III studies included 525 cases and 512 controls matched for age and for the place of recruitment; the invasive-cancer studies included 373 histologically confirmed cases of squamous-cell carcinoma and 387 age-stratified controls selected randomly from the populations that generated the cases. Antibodies to Chlamydia trachomatis, Neisseria gonorrhoeae, Treponema pallidum, herpes simplex virus type II (HSV-2) and cytomegalovirus (CMV) were tested in 88% of the women. Cervical scrapes were examined for HPV DNA in 63% of the women using a polymerase-chain-reaction assay (PCR). Among controls, the highest antibody prevalence was to CMV (96.5%), followed by HSV-2 (31.4%) and C. trachomatis (23.3%). For all STAs, the sero-prevalence was markedly higher in Colombia than in Spain both for cases and for controls. After adjustment for the presence of HPV DNA, C. trachomatis was the only STA associated with CIN III in both countries; Spain and Colombia. In both countries, the risk of CIN III increased with increasing of C. trachomatis antibody titers. Among Spanish women, an increase in risk of invasive carcinoma was found for those with antibodies to N. gonorrhoeae; those with antibodies to HSV-2 and those with antibodies to C. trachomatis. These associations were present only in HPV-DNA-negative women. Among HPV-DNA-positive women, none of the STAs considered were associated with cervical neoplasia. Our findings could be interpreted as indicating that past infections with HSV-2, N. gonorrhoeae and C. trachomatis are surrogate markers of HPV, but because HPV DNA may have escaped detection, we cannot exclude that these STAs are also of separate etiological significance.

Case-Control Studies↗
PubMed1994

Humoral immune response to plasmid protein pgp3 in patients with Chlamydia trachomatis infection.

We identified, by two-dimensional electrophoretic analysis and microsequencing, a protein of Chlamydia trachomatis elementary bodies which corresponds to the polypeptide (pgp3) encoded by open reading frame 3 (ORF3). Amino acid analysis showed that the first residue (Gly) found in the native protein is the one encoded by the second ORF3 codon, implying a typical bacterial removal of the first Met residue. Relatively large amounts of recombinant pgp3 (r-pgp3) in a stable, water-soluble form were obtained by overexpressing ORF3 in Escherichia coli and purifying the product from periplasmic extracts under nondenaturing conditions. These r-pgp3 preparations allowed specific detection of anti-pgp3 antibodies by enzyme-linked immunosorbent assay. Analysis of a group of 170 sera from healthy blood donors and from patients who were seropositive or -negative for C. trachomatis and Chlamydia pneumoniae showed that an immune response to pgp3 occurs in the majority (ca. 81%) of patients with sexually transmitted diseases who are seropositive for C. trachomatis and generally correlates with the response to cell surface antigens. No reaction between r-pgp3 and C. pneumoniae-positive sera was detected.

Amino Acid Sequence↗
PubMed1994

[Serodiagnosis of Chlamydia trachomatis and Chlamydia pneumoniae. Evaluation comparing 2 methods: microimmunofluorescence and ELISA. A study of 216 Congolese sera].

Our purpose was to compare two assays for C. trachomatis and C. pneumoniae serology: immunofluorescence and ELISA. With both assays, we tested 216 sera from two populations: risk population of STD (sexually transmitted disease) (women) and a control population "blood donors". Positives percentages of each population were compared by two assays: 87% in MIF and 76% in ELISA (women population); 64.8% in MIF and 53.7% in ELISA (blood donors population). Results were not significantly different. In comparison with ELISA: MIF had a sensitivity of 85% and a specificity of 93%. In comparison with MIF: ELISA had a sensitivity of 98% and a specificity of 48%; agreement between MIF and ELISA in women population and blood donors population is respectively: 88.9% and 78.8%. The discrepancy between both methods is 11.1% in women group and 21.2% in blood donors group. The correlated coefficient (r) between MIF and ELISA was ranged between 0.507 and 0.778 (P < 0.001). Both assays have honourable performance, but no superimposable.

Adolescent↗