Development of aChlamydia suis-specific antibody enzyme-linked immunosorbent assay based on the use of a B-cell epitope of the polymorphic membrane protein C

Abstract

Chlamydia suis infections lead to economic loss in the pork industry. Chlamydia suis infections could be successfully treated with tetracyclines until the appearance of a tetracycline resistant phenotype, which was acquired via horizontal gene transfer of the tet(C) gene. Given the importance of C. suis as a swine pathogen and as a recently emerged tetracycline resistant pathogen with zoonotic potential, our aim was to develop a sensitive C. suis-specific antibody ELISA based on the polymorphic membrane proteins (Pmps). Chlamydia Pmps are important virulence factors and candidate antigens for serodiagnosis. We identified nine Pmps (PmpA to I) in C. suis strain MD56 using a recently developed Hidden-Markov model. PmpC was the most promising candidate for the development of a C. suis-specific antibody ELISA as the protein was absent in C. abortus, C. pecorum and C. psittaci which also infect pigs and as the protein contained C. suis-specific amino acid regions, absent in C. trachomatis PmpC. We identified an immunodominant B-cell epitope in C. suis PmpC using experimental porcine sera. The sensitivity and specificity of the PmpC ELISA was compared to the complement fixation test (CFT) and to a recombinant MOMP ELISA using experimental sera. The PmpC ELISA detected all positive control sera and was in contrast to CFT and the rMOMP ELISA 100% C. suis specific as positive control sera against other Chlamydia species did not react in the PmpC ELISA. The test was successfully validated using slaughterhouse sera and sera from clinically affected pigs. The PmpC ELISA could assist in diminishing the spread of C. suis infections in the pork industry.

LA PREDICCIÓN IN SILICO DETERMINA LA PRESENCIA DE EPÍTOPOS INMUNOGÉNICOS ALTAMENTE ESPECÍFICOS EN FRAGMENTOS DE LA PROTEÍNA DE LA MEMBRANA POLIMÓRFICA (PMP17G) DE Chlamydia abortus

“…Aunque en la mayoría de los casos este tipo de herramientas se han empleado para el desarrollo de vacunas recombinantes (Soria-Guerra et al, 2015;Aguilar-Montes de Oca et al, 2022); en el presente estudio y otros realizados previamente, se ha demostrado que estos también pueden emplearse para el desarrollo de pruebas de serodiagnóstico, debido a la naturaleza de las mismas; por lo cual, es posible emplearlas para dicho fin como ya se ha llevado a cabo en el caso de C. suis (De Puysseleyr et al, 2018).…”

Characteristics of Chlamydia suis Ocular Infection in Pigs

“…Sera were analysed using different methods for antibody detection: CFT [ 43 ], an in-house recombinant MOMP ELISA [ 19 , 30 ], and a PmpC-based ELISA [ 31 ]. Full-length recombinant MOMP of C. suis S45 was produced in COS-7 cells as previously described [ 44 ] and used to coat ELISA plates.…”

“…Pathogens 2021, 10, x FOR PEER REVIEW 3 of 14 C. suis-specific polymorphic outer membrane protein C (PmpC)-based ELISA [31] in piglets over three weeks after infection. A potential recognition of C. suis antigens by T cells was investigated by IFN-γ ELISpot assays.…”

“…Our study aimed to evaluate the distribution of C. suis S45 in organs besides conjunctiva after a primary experimental conjunctival infection and to measure levels of C. suis -specific antibodies using (i) the complement fixation test, (ii) an in-house Chlamydiaceae -specific major outer membrane protein (MOMP) ELISA [ 19 , 30 ], and (iii) a C. suis -specific polymorphic outer membrane protein C (PmpC)-based ELISA [ 31 ] in piglets over three weeks after infection. A potential recognition of C. suis antigens by T cells was investigated by IFN-γ ELISpot assays.…”

Silver Nanocatalyst Based Clock Reaction for Multi‐mode Detection of Tetracycline Antibiotics

“…[1][2][3][4] However, the residual problem of TCs and their metabolites in foodstuffs resulting from the drug abuse will cause seriously bad effects such as hepatotoxicity, gastrointestinal disorders, allergic reactions and drug resistance. [5][6][7] A number of analytical methods such as fluorescence, [8][9][10] high performance liquid chromatography tandem-mass spectrometry (HPLC-MS), [11][12][13][14] electrophoresis, [15][16] immunochromatography, [17,18] and enzyme linked immunosorbent assay [19][20][21][22] have been well developed for monitoring the levels of TCs residues, however, these reported methods always require expensive instruments and sophisticated operating procedures. Hence, the development of convenient and cost-effective assay for TCs is highly desirable.…”