A novel synthetic peptide microarray assay detects Chlamydia species-specific antibodies in animal and human sera

Abstract

Serological analysis of Chlamydia (C.) spp. infections is still mainly based on micro-immunofluorescence and ELISA. To overcome the limitations of conventional serology, we have designed a novel microarray carrying 52 synthetic peptides representing B-cell epitopes from immunodominant proteins of all 11 chlamydial species. The new assay has been validated using monospecific mouse hyperimmune sera. Subsequently, serum samples from cattle, sheep and humans with a known history of chlamydial infection were examined. For instance, the specific humoral response of sheep to treatment with a C. abortus vaccine has been visualized against a background of C. pecorum carriership. In samples from humans, dual infection with C. trachomatis and C. pneumoniae could be demonstrated. The experiments revealed that the peptide microarray assay was capable of simultaneously identifying specific antibodies to each Chlamydia spp. The actual assay represents an open platform test that can be complemented through future advances in Chlamydia proteome research. The concept of the highly parallel multi-antigen microarray proven in this study has the potential to enhance our understanding of antibody responses by defining not only a single quantitative response, but also the pattern of this response. The added value of using peptide antigens will consist in unprecedented serodiagnostic specificity.

Discovery of Human-Specific Immunodominant Chlamydia trachomatis B Cell Epitopes

“…These synthetic peptide antigens can be readily commercialized in the standard ELISA format as well as in the microarray format ( 63 , 66 , 67 ), and such assays may become widely available tools for chlamydial serology. In a recent study ( 66 ), we detected species- and serovar-specific antibodies against Chlamydia spp. by a peptide microarray that used several of the peptide antigens described here ( Fig.…”

Identification of a Universal Antigen Epitope of Influenza A Virus using Peptide Microarray

“…In a dot matrix, proteins are decomposed into a plurality of peptides and dotted on silica gel lm [33,34]. It can not only detect the corresponding antibody in serum but also reverse deduce the epitope recognized by the antibody [35,36]. Based on the peptide microarray, we screened out the 14 th peptide, KELGNGCFEFYHKCDNECME in HA2, which was positively responsive to the antibodies against multiple subtypes of IAVs.…”

Use of meat juice and blood serum with a miniaturised protein microarray assay to develop a multi-parameter IgG screening test with high sample throughput potential for slaughtering pigs

“…To overcome this obstacle, we previously reported the development of a miniaturised protein microarray as a new serological IgG screening test for zoonotic agents and production diseases in pigs [5]. Protein microarrays are excellently suited for the simultaneous detection of different analytes and have already been used for the simultaneous detection of different antibodies [6][7][8]. By coupling different antigens on the microarray chip surface, the respective antibodies can be detected in a joint test run, which not only saves costs, but also analysis time.…”