Characterization of a monoclonal antibody to the group antigen of Chlamydia spp. and its use for antigen detection by reverse passive haemagglutination and indirect immunofluorescence

Thornley, Margaret J.

doi:10.1016/0378-1097(83)90224-0

Cited by 3 publications

(11 citation statements)

References 7 publications

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“…IMS uses antibodies associated with magnetic beads to bind intact Chlamydia, with wash steps designed to remove contaminating material, enabling enrichment of the target species. We used a commercially available anti-Chlamydia mouse IgG primary antibody (IMAGEN Chlamydia, Oxoid) against C. trachomatis lipopolysaccharide (LPS), which binds to all serovars of C. trachomatis and has been tested for cross-reactivity against many other microbial species including Lactobacillus lactis, Mycoplasma spp., Neisseria gonorrhoeae, and Gardnerella vaginalis (IMAGEN Chlamydia booklet; Thornley et al 1983Thornley et al , 1985. LPS is present at ;34,000 molecules per EB (Su et al 1990), creating a high density target for antibody binding.…”

Section: Developing Ims-mda For C Trachomatismentioning

confidence: 99%

Whole-genome sequences ofChlamydia trachomatisdirectly from clinical samples without culture

et al. 2013

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The use of whole-genome sequencing as a tool for the study of infectious bacteria is of growing clinical interest. Chlamydia trachomatis is responsible for sexually transmitted infections and the blinding disease trachoma, which affect hundreds of millions of people worldwide. Recombination is widespread within the genome of C. trachomatis, thus whole-genome sequencing is necessary to understand the evolution, diversity, and epidemiology of this pathogen. Culture of C. trachomatis has, until now, been a prerequisite to obtain DNA for whole-genome sequencing; however, as C. trachomatis is an obligate intracellular pathogen, this procedure is technically demanding and time consuming. Discarded clinical samples represent a large resource for sequencing the genomes of pathogens, yet clinical swabs frequently contain very low levels of C. trachomatis DNA and large amounts of contaminating microbial and human DNA. To determine whether it is possible to obtain whole-genome sequences from bacteria without the need for culture, we have devised an approach that combines immunomagnetic separation (IMS) for targeted bacterial enrichment with multiple displacement amplification (MDA) for whole-genome amplification. Using IMS-MDA in conjunction with high-throughput multiplexed Illumina sequencing, we have produced the first whole bacterial genome sequences direct from clinical samples. We also show that this method can be used to generate genome data from nonviable archived samples. This method will prove a useful tool in answering questions relating to the biology of many difficult-to-culture or fastidious bacteria of clinical concern.

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Section: Developing Ims-mda For C Trachomatismentioning

confidence: 99%

Whole-genome sequences ofChlamydia trachomatisdirectly from clinical samples without culture

et al. 2013

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“…Monoclonal antibodies to the chlamydial genus-specific antigen have been described by Stephens et al (1982) and Thornley et al (1983). Our study of the production and charalcteristics of such monoclonal antibodies, and their possible eventual application in diagnostic methods (Thornley et al, 1983) has been extended to include four additional monoclonal antibodies, characterization of their reactions with chlamydial antigens, a study of cross-reactions with Salmonella Re-LPS and Acinetobacter strains, and further quantitative characterization of the chlamydial genus-specific antigen.…”

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confidence: 90%

“…trachomatis strains 48/80H and SA2(f) were cultured in McCoy cells (Evans & Taylor-Robinson, 1979); SA2(f) was additionally cultured in egg yolk sacs (Schachter & Dawson, 1978). Elementary bodies from egg yolk cultures were purified by density centrifugations and enzyme treatment, followed by extraction with 1 % (w/v) sodium deoxycholate (DOC) to yield the genusspecific antigen (Thornley et a/., 1983).…”

Section: E T H O D Smentioning

confidence: 99%

“…Our study of the production and charalcteristics of such monoclonal antibodies, and their possible eventual application in diagnostic methods (Thornley et al, 1983) has been extended to include four additional monoclonal antibodies, characterization of their reactions with chlamydial antigens, a study of cross-reactions with Salmonella Re-LPS and Acinetobacter strains, and further quantitative characterization of the chlamydial genus-specific antigen. The sensitivity of the monoclonal antibodies when coupled to trypsin-treated sheep red blood cells and then used to detect chlamydial antigen b:y reverse passive haemagglutination (RPH) has also been studied.…”

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confidence: 99%

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Properties of Monoclonal Antibodies to the Genus-specific Antigen of Chlamydia and Their Use for Antigen Detection by Reverse Passive Haemagglutination

et al. 1985

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The chlamydial genus-specific antigen was extracted with phenol/chloroform/petroleum ether (PCP) from preparations of Chlamydia trachomatis and C. psittaci, and quantities measured using an assay for lipopolysaccharide (LPS). The LPS from C. trachomatis contained 2.2% (w/w) of ketodeoxyoctanoic acid. Five IgG monoclonal antibodies reacted in an ELISA with LPS from both species, the antigen being periodate-sensitive and heat-resistant, confirming that all antibodies were against the genus-specific antigen. All the antibodies bound to the PCP extract of C. trachomatis on an immunoblot, at a position corresponding to the periodate-Schiff-stained bands of both C. trachomatis extract and Salmonella Re-LPS. When linked to trypsin-treated sheep erthrocytes and used in reverse passive haemagglutination tests, all antibodies gave indicator cells capable of detecting chlamydial LPS or crude preparations of chlamydiae grown in McCoy cells, the sensitivity varying with the antibody used. The antibodies varied in IgG subclass (either IgG2a or IgG3), and in ability to precipitate in immunodiffusion tests. Two antibodies cross-reacted with one strain of Acinetobacter in ELISA and with Salmonella Re-LPS in both ELISA and immunodiffusion tests. The other three did not react in ELISA with Acinetobacter strains or Salmonella Re-LPS, and none of the five reacted with LPS of E. coli or Pseudomonas morsprunorum.

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“…Blood was collected two to three weeks after the last injection. ELISA was done essentially as described by Thornley et al (1983) except that the polystyrene microtitre plates (Nunc-Immuno Plate 11) were coated by filling the wells of the plate with 50 pl LPS (5 pg ml-I) in 0.1 M-Na2C03 buffer, pH 9.6, and incubating for 18 h at 4 "C. Bound antibodies were detected with a peroxidase-conjugated anti-mouse immunoglobulin serum (Dako). o-Phenylenediamine (OPD) was used as the substrate for the enzyme and absorbance was measured at 492 nm.…”

Section: Haemophilus Influenzae Lipopolysaccharide 1445mentioning

confidence: 99%

Composition of the Lipopolysaccharide from Different Capsular Serotype Strains of Haemophilus influenzae

Se¹,

Moxon²

1987

Microbiology

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Lipopolysaccharide (LPS) from all six serotype strains of Haemophilus injhenzae was similar in composition. The oligosaccharide, of each LPS, was composed of glucose, galactose, heptose and 2-keto-3-deoxyoctonic acid. The lipid A was composed of glucosamine, phosphate and the fatty acids 14:O and 3-OH 14:O. Each LPS also contained ethanolamine and ethanolamine phosphate, and the oligosaccharides from two strains additionally contained small amounts of glucosamine. Although the LPS was similar in composition, different serotypes had quantitative differences, especially in the galactose content, which correlated with the antigenic specificity of their homologous antisera and with their mobility on SDS-polyacrylamide gel electrophoresis (SDS-PAGE). A survey by SDS-PAGE showed that LPS from strains of the serotypes a, c and d was characteristically of lower M , than the LPS from most (80%) serotype b strains. I N T R O D U C T I O NHaemophilus inzuenzae is widespread in the human population, where it colonizes the nasopharynx. Although normally commensal, it has the potential to cause systemic infection in children, including meningitis. It is either non-capsular or expresses one of six antigenically and chemically distinct capsular polysaccharides, designated types a-f (Pittman, 193 1). The majority of strains causing systemic infection synthesize the type b capsule, an acidic polysaccharide with the repeating unit of ribosyl-ribitol-phosphate (PRP) (Moxon & Vaughn, 1981; Turk, 1982). Although the type b capsule is known to be a major virulence factor, a potential role for non-capsular surface components such as the lipopolysaccharide (LPS) of this organism has been indicated by the results of recent studies. These have shown an association between virulence and alterations in LPS as a result of transformation (Zwahlen et al., 1983(Zwahlen et al., , 1985 and also an apparent relationship between virulence and the presence of certain antigenic determinants in H. inpuenzae type b LPS as defined by monoclonal antibodies (Kimura & Hansen, 1986).H. influenzae LPS also displays endotoxic properties similar to those of enterobacterial LPS (Flesher & Insel, 1978) and may be involved in the acquisition of serum resistance . Previous studies have shown the LPS to possess a lipid A moiety and an oligosaccharide, but to lack a high-M, sidechain polysaccharide (Flesher & Insel, 1978;Inzana, 1983). Because of the apparent rough nature of this LPS the term lipo-oligosaccharide is currently being employed by a few authors. However, we prefer to retain the original terminology in order to specify a molecule which has the biological and structural characteristics of an LPS.

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Characterization of a monoclonal antibody to the group antigen of Chlamydia spp. and its use for antigen detection by reverse passive haemagglutination and indirect immunofluorescence

Cited by 3 publications

References 7 publications

Whole-genome sequences ofChlamydia trachomatisdirectly from clinical samples without culture

Whole-genome sequences ofChlamydia trachomatisdirectly from clinical samples without culture

Properties of Monoclonal Antibodies to the Genus-specific Antigen of Chlamydia and Their Use for Antigen Detection by Reverse Passive Haemagglutination

Composition of the Lipopolysaccharide from Different Capsular Serotype Strains of Haemophilus influenzae

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