Detection of Chlamydia trachomatis-specific antibodies in human sera by recombinant major outer-membrane protein polyantigens

Mygind, Per; Christiansen, Gunna; Persson, Kenneth; Birkelund, Svend

doi:10.1099/0022-1317-49-5-457

Cited by 16 publications

(17 citation statements)

References 13 publications

(20 reference statements)

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“…The assay that had the best correlation with WIF, particularly at high WIF titres, was the Genzyme Virotech C trachomatis EIA, which uses C trachomatis LGV type II strain, cultured in mouse L cells, and inactivated using γ irradiation. It is possible that the reduced correlation of the synthetic peptide assays results from conformational differences in the epitopes presented by the peptides 14 compared with native antigens.…”

Section: Discussionmentioning

confidence: 99%

Measurement of IgG antibodies to Chlamydia trachomatis by commercial enzyme immunoassays and immunofluorescence in sera from pregnant women and patients with infertility, pelvic inflammatory disease, ectopic pregnancy, and laboratory diagnosed Chlamydia psittaci/Chlamydia pneumoniae infection

Jones¹

2003

Journal of Clinical Pathology

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Background: Screening for Chlamydia trachomatis specific antibodies is valuable in diagnosing asymptomatic pelvic inflammatory disease (PID) and tubal damage following repeated episodes of PID. The assays in current use are unsuitable for screening large numbers of samples so there is a need to develop more suitable assays. Aims: To compare the performance of several commercial C trachomatis enzyme immunoassays (EIAs) (SeroCT, C tracho pep , Medac p-EIA, Vircell and Labsystems C trachomatis IgG EIAs) using major outer membrane protein (MOMP), an inactivated organism EIA (Genzyme Virotech EIA), and a genus specific EIA (Platelia Chlamydia IgG) with the whole cell inclusion immunofluorescence (WIF) assay. In addition, to adapt, using time resolved fluorescence technology, the assay showing the highest correlation with WIF. Methods: Ninety sera from patients presenting with ectopic pregnancies, 187 sera from those with a variety of types of infertility, 33 sera from cases of PID where a fourfold rise in WIF titre occurred, and 90 sera from antenatal clinic attenders were tested. A panel of 36 sera from laboratory diagnosed cases of Chlamydia psittaci/Chlamydia pneumoniae infection was also tested. Results: The Genzyme Virotech EIA showed the highest rank correlation coefficient (0.82) with WIF, particularly at high WIF titres. The MOMP specific assays varied in their correlation with WIF, with rank correlation coefficients ranging from 0.70 (Medac p-EIA) to 0.80 (Vircell EIA). The Genzyme Virotech assay showed poor specificity (5.6%; 95% confidence interval (CI), 0.68% to 18.7%)-it was reactive with 34 of the panel of 36 C psittaci/ C pneumoniae positive sera. The MOMP based EIAs showed high specificity, particularly the Medac p-ELISA (97.2%; 95% CI, 85.5% to 99.9%)-only one serum was reactive. In view of the good correlation between WIF and the Genzyme Virotech EIA, a time resolved fluorescence immunoassay (TRFIA) was developed using the Genzyme Virotech antigen. Using an appropriate cut off the TRFIA assay showed excellent correlation with WIF. Conclusions: The TRFIA assay may be useful as a screening assay, possibly in conjunction with one of the highly specific EIAs studied (for example, Medac p-EIA) to confirm the antibody specificity of sera selected by the screening assay.

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Section: Discussionmentioning

confidence: 99%

Measurement of IgG antibodies to Chlamydia trachomatis by commercial enzyme immunoassays and immunofluorescence in sera from pregnant women and patients with infertility, pelvic inflammatory disease, ectopic pregnancy, and laboratory diagnosed Chlamydia psittaci/Chlamydia pneumoniae infection

Jones¹

2003

Journal of Clinical Pathology

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“…Cloning and expression of porB was done using the pET-30 LIC Vector Kit (Novagen, Madison, WI, USA) according to the manufacturer's instructions. The histidine-tagged PorB fusion protein was purified by affinity chromatography (Quagen, Valencia, CA, USA) under denaturing conditions as described in [25] using a nickel column (High Trap Sepharose, Amersham Pharmacia Biotech). The correct PorB sequence was confirmed by analyzing the fusion protein excised from a Commasie stained 1-D SDS-PAGE gel by MALDI-MS. Sera containing polyclonal antibodies reacting with PorB (PAb 242) was obtained from a rabbit that was immunized with purified PorB fusion protein.…”

Section: Cloning and Expression Of Porb And Generation Of Antibodies mentioning

confidence: 99%

Comparative proteome analysis ofChlamydia trachomatis serovar A, D and L2

et al. 2002

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Chlamydia trachomatis represents a group of human pathogenic obligate intracellular and gram-negative bacteria. The genome of C. trachomatis D comprises 894 open reading frames (ORFs). In this study the global expression of genes in C. trachomatis A, D and L2, which are responsible for different chlamydial diseases, was investigated using a proteomics approach. Based on silver stained two-dimensional polyacrylamide gel electrophoresis (2-D PAGE), gels with purified elementary bodies (EB) and auto-radiography of gels with 35S-labeled C. trachomatis proteins up to 700 protein spots were detectable within the range of the immobilized pH gradient (IPG) system used. Using mass spectrometry and N-terminal sequencing followed by database searching we identified 250 C. trachomatis proteins from purified EB of which 144 were derived from different genes representing 16% of the ORFs predicted from the C. trachomatis D genome and the 7.5 kb C. trachomatis plasmid. Important findings include identification of proteins from the type III secretion apparatus, enzymes from the central metabolism and confirmation of expression of 25 hypothetical ORFs and five polymorphic membrane proteins. Comparison of serovars generated novel data on genetic variability as indicated by electrophoretic variation and potentially important examples of serovar specific differences in protein abundance. The availability of the complete genome made it feasible to map and to identify proteins of C. trachomatis on a large scale and the integration of our data in a 2-D PAGE database will create a basis for post genomic research, important for the understanding of chlamydial development and pathogenesis.

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“…However, there is some controversy on whether all 9 of the C. trachomatis Pmps are expressed. Early investigations detected only PmpB, D, E, F, G and H, [50][51][52] whereas later studies demonstrated that all C. trachomatis pmp genes were transcribed. 17,53 C. trachomatis pmp genes are located in 2 clusters with pmpA-C and pmpE-I comprising each cluster and pmpD being genetically isolated.…”

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confidence: 97%

Chlamydial polymorphic membrane proteins: regulation, function and potential vaccine candidates

et al. 2015

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Detection of Chlamydia trachomatis-specific antibodies in human sera by recombinant major outer-membrane protein polyantigens

Cited by 16 publications

References 13 publications

Measurement of IgG antibodies to Chlamydia trachomatis by commercial enzyme immunoassays and immunofluorescence in sera from pregnant women and patients with infertility, pelvic inflammatory disease, ectopic pregnancy, and laboratory diagnosed Chlamydia psittaci/Chlamydia pneumoniae infection

Measurement of IgG antibodies to Chlamydia trachomatis by commercial enzyme immunoassays and immunofluorescence in sera from pregnant women and patients with infertility, pelvic inflammatory disease, ectopic pregnancy, and laboratory diagnosed Chlamydia psittaci/Chlamydia pneumoniae infection

Comparative proteome analysis ofChlamydia trachomatis serovar A, D and L2

Chlamydial polymorphic membrane proteins: regulation, function and potential vaccine candidates

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