Mycobacterial Antigenic Extracts; A View of Their Characterizations And Diagnostic Efficacy

Abstract

With the procedure described in this work, it is possible to prepare and use successfully M. bovis cell extract antigens "sonicated antigen (SA) & membrane associated antigen (MAA)" and culture filtrate antigens "(HK-CF), (ST-CF) and (SU-CF)" in vitro for diagnosis ofbovine TB in comparison with bovine PPD by measuring cell mediated immunity using skin sensitivity test, also by measuring antibodies using ELISA test.

SDS-PAGE was applied for characterization of antigenic extracts and

CF of M. bovis. SA revealed proteins at Mw of 103.8, 64, 53.9, 47.8, 45, 38,

35, 32, 30, 27, 21.4, 15.2, 12.6 and 9.6kDa, MAA revealed proteins at Mw of 103.7, 64, 55.6, 48.5, 45, 38, 35, 32, 30, 26.3, 23.5, 21.4, 16, 12.6 and 9.6kDa, HK-CF revealed proteins at Mw of 48.5, 45, 38, 35, 33, 30.7, 26.3,

21.4, 19, 18.2 and 13, 9.6kDa, ST-CFrevealed proteins at Mw of66.2, 57.6,

52.6, 49, 41.3, 38, 35, 33, 30, 27, 23.5, 21.4, 20.5, 18.2, 14.6, ll.5 and

9.6kDa, SU-CFrevealed proteins at Mw of 57.6, 48.5, 40, 36, 35, 32, 30, 29,

23, 22.4, 19.6, 18.1, 17.4, 14.6, 12.6, 10.9 and 9.6kDa & PPD revealed proteins at Mw of 38.6, 36, 34, 30, 28, 25, 22.1, 16.1, 13.3, 10 and 9.3kDa.

Western blotting analysis was performed, clear visible immunoreactive bands in SA at Mw of 103.7, 82.7, 61.2, 33 and 31.3kDa, in MAA at 121.8, 103.7, 94.5, 82.7, 66.2, 33, 32 and 31kDa, in HK-CF at 124.4, 103.7, 85.2,

66.2, 35 and 33kDa, in ST-CF at 102.2, 82.7 and 57.2kDa, in SU-CF at

102.2, 85.2 and 57.2kDa, and in PPD at 135.2, 117.2 and 85.2kDa.