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. 2023 Feb 20;13(1):2936.
doi: 10.1038/s41598-023-30147-4.

Differences in skin test reactions to official and defined antigens in guinea pigs exposed to non-tuberculous and tuberculous bacteria

Leire Fernández-Veiga  1 , Miguel Fuertes  1 , María V Geijo  1 , Bernat Pérez de Val  2   3 , Enric Vidal  2   3 , Lorraine Michelet  4 , María Laura Boschiroli  4 , Alberto Gómez-Buendía  5   6 , Javier Bezos  5   6 , Gareth J Jones  7 , Martin Vordermeier  7 , Ramón A Juste  1 , Joseba M Garrido  1 , Iker A Sevilla  8
Affiliations

Differences in skin test reactions to official and defined antigens in guinea pigs exposed to non-tuberculous and tuberculous bacteria

Leire Fernández-Veiga et al. Sci Rep. .

Abstract

The single and comparative intradermal tuberculin tests (SITT and CITT) are official in vivo tests for bovine tuberculosis (TB) diagnosis using bovine and avian purified protein derivatives (PPD-B and PPD-A). Infection with bacteria other than Mycobacterium tuberculosis complex (MTC) can result in nonspecific reactions to these tests. We evaluated the performance of the skin test with PPDs and new defined antigens in the guinea pig model. A standard dose (SD) of Rhodococcus equi, Nocardia sp., M. nonchromogenicum, M. monacense, M. intracellulare, M. avium subsp. paratuberculosis, M. avium subsp. avium, M. avium subsp. hominissuis, M. scrofulaceum, M. persicum, M. microti, M. caprae and M. bovis, and a higher dose (HD) of M. nonchromogenicum, M. monacense, M. intracellulare, M. avium subsp. paratuberculosis were tested using PPD-B, PPD-A, P22, ESAT-6-CFP-10-Rv3615c peptide cocktail long (PCL) and fusion protein (FP). The SD of R. equi, Nocardia sp., M. nonchromogenicum, M. monacense, M. intracellulare and M. avium subsp. paratuberculosis did not cause any reactions. The HD of M. nonchromogenicum, M. monacense, M. intracellulare, and M. avium subsp. paratuberculosis and the SD of M. avium subsp. hominissuis, M. scrofulaceum and M. persicum, caused nonspecific reactions (SIT). A CITT interpretation would have considered M. avium complex and M. scrofulaceum groups negative, but not all individuals from M. nonchromogenicum HD, M. monacense HD and M. persicum SD groups. Only animals exposed to M. bovis and M. caprae reacted to PCL and FP. These results support the advantage of complementing or replacing PPD-B to improve specificity without losing sensitivity.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any competing interests. G.J.J. and H.M.V. are employed by the Animal and Plant Health Agency (APHA) who holds 3 patents for the use of Rv3615c in diagnostic tests for bovine TB (WO/2009/060184, WO/2011/135369 and WO/2012/010875).

Figures

Figure 1
Figure 1
Reactivity according to inoculated organism and test antigen. Individual (I, II and III) and mean (M) erythema areas read 24 h after skin test antigens inoculation are plotted. The panel at the left on the top serves to represent the control group that was not inoculated any organisms (Ø), and the standard dose (SD) of Rhodococcus equi (Rho), Nocardia sp. (Noc), M. nonchromogenicum (Mno), M. monacense (Mmo), M. intracellulare (Min) and M. avium subsp. paratuberculosis (Map). The rest of results correspond to the SD of the organism indicated except for M. nonchromogenicum, M. monacense, M. intracellulare and M. avium subsp. paratuberculosis (M. avium subsp. paratb) that correspond to inoculations with the higher dose (HD). B‒A indicates the result of subtracting PPD-A result to PPD-B result (PPD-B‒PPD-A).
Figure 2
Figure 2
Effect of the time of reading (24 or 48 h) on the erythematous area according to inoculated organism and intradermal antigen. Results refer to inoculations with the standard dose (SD) except for M. nonchromogenicum, M. monacense, M. intracellulare and M. avium subsp. paratuberculosis that correspond to inoculations with the higher dose (HD).
Figure 3
Figure 3
Position of skin test antigens. Site of injection of skin test antigens in guinea pigs according to a Latin square arrangement considering each side of each animal as independent.
See this image and copyright information in PMC

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