Table 1.
Borrelia burgdorferi senso stricto-specific antigens applied on nitrocellulose strips of three LIAs used.
Table 2.
LIA A–overall rating of a test strip or equine serum sample and its assignment to an overall test result allocation category according to manufacturers’ instructions.
Table 3.
LIA B–overall rating of a test strip or equine serum sample and its assignment to an overall test result allocation category according to manufacturers’ instructions.
Table 4.
LIA C–overall rating of a test strip or equine serum sample and its assignment to an overall test result allocation category according to manufacturers’ instructions.
Fig 1.
(A)-(I). Comparison of test results obtained by three LIAs used for Bbsl-serodiagnosis with equine serum samples. Evaluation was performed according to the three manufacturers’ instruction manuals. Test results are charted by experimental group (Non-Vac (n = 42), Vac-Basic (n = 45), Vac-Plus (n = 44)), LIA (LIA A, LIA B and LIA C) and time-point of blood sample collection (d0, d135, d210). (A)-(C). Comparison of OspA AG-AB immunocomplex signal intensity results. The number of serum samples with a “negative”OspA signal intensity result are displayed in white (no signal) and lilac (signal fainter than COC), the number of samples with a “positive”OspA signal intensity result are illustrated in light purple (signal equal to COC) and bright purple (signal greater than COC). (D)-(F). Comparison of VlsE AG-AB immunocomplex signal intensity results. The number of serum samples with a “negative”VlsE signal intensity result are represented by white color (no signal) and vanilla (signal fainter than COC), number of samples with a “positive”VlsE signal intensity result are displayed in yellow (signal equal to COC) and ocher (signal greater than COC). (G)-(I). Comparison of overall test results. The number of serum samples with an overall test result of “NEGATIVE” (green), “EQUIVOCAL”(salmon), “POSITIVE (infection)”(red), “VACCINATED AND POSITIVE” (light brown) and “VACCINATED” (blue) are illustrated. In LIA A, vaccination-specific OspA is not considered as evidence for immunization and, therefore, there is no allocation in terms of “VACCINATED” and “VACCINATED AND POSITIVE” in this test. Bbsl, Borrelia burgdorferi sensu lato; Non-Vac, non-vaccinated horses; Vac-Basic, horses vaccinated on d0 and d14; Vac-Plus, horses vaccinated on d0, d14 and d180; d, day of blood sample collection during the vaccination schedule; AG, antigen; AB, antibody; OspA, outer surface protein A; VlsE, variable major protein-like sequence expressed; COC, cut off control.
Table 5.
Recommended overall evaluation protocol (ROEP)–overall rating of a test strip or equine serum sample and its assignment to an overall test result allocation category according the ROEP.
Fig 2.
(A)-(C). Overall test results after application of the ROEP–comparison of overall test results obtained by three LIAs used for LB serodiagnosis in horses. Evaluation and allocation were performed according to an alternative recommended overall evaluation protocol for equine serum samples which considered OspA signal as specific for equine immunoreaction. The number of serum samples with an overall test result of “NEGATIVE” (green), “EQUIVOCAL”(salmon), “POSITIVE”(red), “VACCINATED AND POSITIVE” (light brown) and “VACCINATED” (blue) after evaluation with the ROEP are illustrated. Results are charted by experimental group (Non-Vac (n = 42), Vac-Basic (n = 45), Vac-Plus (n = 44)), LIA (LIA A, LIA B, LIA C) and time-point of blood sample collection (d0, d135 and d210). ROEP, recommended overall evaluation protocol; d, day of blood sample collection during the vaccination schedule; Non-Vac, non-vaccinated horses; Vac-Basic, horses vaccinated on d0 and d14; Vac-Plus, horses vaccinated on d0, d14 and d180.