Comparison of three commercial serological tests for the detection of Chlamydia abortus infection in ewes

Abstract

Background: 20.9% of diagnosable abortions in Ireland in 2015 were caused by Chlamydia abortus infection. Abortion usually occurs in the last 2-3 weeks of gestation, and up to 30% of ewes may be affected in naïve flocks. Serological diagnosis of EAE in flocks using LPS or whole bacteria as antigens is often hindered by cross reactions with C. pecorum. Although the complement fixation test is the official test for diagnosis of EAE, more sensitive and specific ELISA based tests have been developed. This study aimed to compare three commercial ELISA kits to detect C. abortus antibodies in ewes and to determine which of the kits had the highest sensitivity. The IDvet kit utilises a MOMP peptide antigen, the MVD-Enfer kit is based on a POMP90-3 antigen while the LSI kit plates are coated with chlamydial LPS. The study also aimed to examine the potential of these ELISAs to distinguish infected animals that go on to abort compared to those that have live lambs. Ewes were vaccinated with either a commercial live vaccine (n = 10) or Tris-buffer sham inoculation (n = 9) 5 months prior to gestation, these ewes were then challenged with C. abortus (1 × 106 IFU/ml) on day 90 of gestation. Sera were collected at pre-vaccination, 14 days post vaccination, 35 days post vaccination, pre-challenge, 35 days post challenge and 3 weeks post lambing/abortion (~ 70 days post challenge) and tested using the 3 aforementioned ELISAs to determine if one ELISA was more sensitive at detecting circulating anti-chlamydial antibodies. Results: Sensitivity was highest with the LSI test kit at 94.74%, followed by the MVD-Enfer and IDvet kits, at 78.95 and 73.68% respectively. Ewes vaccinated with Enzovax became seropositive at 14 days post vaccination with all kits. Following challenge at day 90 of gestation, antibody titres steadily rose in all groups of ewes. With all ELISA kits, antibody levels were higher in ewes that aborted compared to ewes that had live lambs at 35 days post challenge and three weeks post lambing, and statistically significantly higher antibody levels were recorded in ewes that aborted compared to ewes that had live lambs using the MVD-ENFER ELISA at three weeks post lambing (P = 0.0482). Conclusions: The LSI assay was the most sensitive out of the three kits tested in this study, when sera were tested at three weeks post lambing. As the LPS used in this kit is cross-reactive with all chlamydia, it is good for identifying flocks infected with any chlamydial species, but it is not considered specific for C. abortus. Furthermore, antibody levels were higher in ewes that aborted compared to ewes that had live lambs, at both 35 days post challenge and at three weeks post lambing. Future work should include evaluation of a larger number of sera at a wider range of time-points as well as an estimation of the specificity of commercially available assays.