Search for Mycobacterium avium Subspecies paratuberculosis Antigens for the Diagnosis of Paratuberculosis

Table 3

Reactivity of bovine sera to the panel of selected antigens from MAP.

Antigen assayed	No. of sera with positive recognition
	control; (n = 10)	PTB infected; (n = 25)	TB infected; (n = 8)
MAP 0011	0	5	4
MAP 0034	5	5	0
MAP 0038	3	8	0
MAP 0047	3	4	0
MAP 0187c	1	1	0
MAP 0209c	4	9	0
MAP 0210c	0	8	0
MAP 0211	0	6	0
MAP 0297	2	4	1
MAP 0334	2	15	1
MAP 0900	0	3	0
MAP 0946	1	8	0
MAP 1012	2	0	0
MAP 1050	4	9	0
MAP 1272	2	8	0
MAP 1293	10	13	1
MAP 1308	2	2	0
MAP 1564	5	1	0
MAP 1589c	2	9	2
MAP 1653	10	20	8
MAP 1693	3	12	2
MAP 1754	0	5	4
MAP 1889c	7	7	3
MAP 1962	3	16	5
MAP 2020	0	9	2
MAP 2167	3	0	0
MAP 2182c	5	1	2
MAP 2513	1	6	2
MAP 2609	2	4	0
MAP 2676	1	4	0
MAP 2685	10	2	0
MAP 2878	10	2	0
MAP 2942	2	7	0
MAP 2956	1	1	1
MAP 3175	4	4	4
MAP 3194	0	1	0
MAP 3205	0	0	0
MAP 3206	1	5	0
Mb 3341	4	6	3
MAP 3402	2	20	1
MAP 3457	10	8	4
MAP 3491	7	7	1
MAP 3527	6	25	8
MAP 3627	3	4	0
MAP 3651	10	14	8
MAP 3692c	5	8	0
MAP 3840	0	3	3
MAP 3841	10	14	1
MAP 3857	10	4	4
MAP 3936	5	8	4
MAP 4000c	3	5	0
MAP 4143	8	16	7
MAP 4147	1	3	0
MAP 4227	1	1	1
PPDA	6	1	4
PPDB	4	16	3
PPA-3	1	18	2
Cocktail	2	14	0

20 uL of antigens were applied to a nitrocellulose membrane (Amersham Hybond-ECL, GE Healthcare Life Sciences, UK) at a concentration of 100 ug/mL, using a semiautomatic aerolizer (Camag Scientific Inc., Wilmington, Delaware). Membranes were evaluated by immunoblotting using sera from healthy and infected animals. Numbers of sera with antibody response are indicated. Samples with intensities values higher than the media obtained with the control (PBS) were considered positive. Antigens selected to evaluate as cocktail are shown in bold. PPA-3: Paratuberculosis protoplasmatic antigen (Allied Monitor, Inc. USA); PPDA and PPDB: avian- and bovine-derivative protein purified (Prionic Switzerland ); Cocktail: Mix of the 7 antigens shown in bold.