Detection of Mycobacterium avium Subspecies paratuberculosis (MAP) Microorganisms Using Antigenic MAP Cell Envelope Proteins

Abstract

Cell envelope proteins from Mycobacterium avium subspecies paratuberculosis (MAP) that are antigenically distinct from closely related mycobacterial species are potentially useful for Johne's Disease (JD) diagnosis. We evaluated the potential of ELISAs, based on six antigenically distinct recombinant MAP cell envelope proteins (SdhA, FadE25_2, FadE3_2, Mkl, DesA2, and hypothetical protein MAP1233) as well as an extract of MAP total cell envelope proteins, to detect antibodies against MAP in the sera of infected cattle. The sensitivity (Se) and specificity (Sp) of an ELISA based on MAP total cell envelope proteins, when analyzing 153 bovine serum samples, was 75 and 96%, respectively. Analysis of the same samples, using a commercial serum ELISA resulted in a Se of 56% and Sp of 99%. Results of ELISA analysis using plates coated with recombinant cell envelope proteins ranged from a highest Se of 94% and a lowest Sp of 79% for Sdh A to a lowest Se of 67% and a highest Sp of 95% for hypothetical protein MAP1233. Using polyclonal antibodies to MAP total cell envelope proteins, immunohistochemical analysis of intestinal and lymph node tissues from JD-positive cattle detected MAP organisms whereas antibodies to recombinant proteins did not. Finally, polyclonal antibodies to MAP total cell envelope protein and to recombinant SdhA, FadE25_2, and DesA2 proteins immunomagnetically separated MAP microorganisms spiked in PBS. These results suggest that antigenically distinct MAP cell envelope proteins and antibodies to these proteins may have potential to detect MAP infection in dairy cattle.

Metabolomic changes in polyunsaturated fatty acids and eicosanoids as diagnostic biomarkers in Mycobacterium avium ssp. paratuberculosis (MAP)-inoculated Holstein–Friesian heifers

“…As a result, they perform poorly between the initial CMI response and the sub (clinical) stages of infection [ 22 ]. The application of omic approaches have suggested that they could be a potential source of new biomarkers with sensitivity and specificity values reported ranging between 82 and 94% [ 11 , 12 ]. However, no omic study has assessed MAP infection associated changes in youngstock.…”

“…These studies suggest that the performance of these omic study-derived biomarkers exceeds many commercially available tests [ 8 ]. This stated, these have focused on post-partum cattle [ 12 , 13 ] so that the application of these biomarkers in youngstock is questionable.…”

“…Omic approaches encapsulate various subdivisions of systems biology, including genomics, proteomics, lipidomics and metabolomics [10]. The application of these approaches in MAP research has highlighted multiple potential biomarkers, but these findings require further validation [11][12][13]. Proteomic approaches identified antigenically distinct recombinant MAP cell envelope proteins; Sdh A and MAP1233m and their detection using ELISA showed maximum sensitivity and specificity values of 94% and 100%, respectively [12].…”

“…The application of these approaches in MAP research has highlighted multiple potential biomarkers, but these findings require further validation [11][12][13]. Proteomic approaches identified antigenically distinct recombinant MAP cell envelope proteins; Sdh A and MAP1233m and their detection using ELISA showed maximum sensitivity and specificity values of 94% and 100%, respectively [12]. Further lipidomic approaches were used to show that glycopeptidolipids are replaced by lipopentapeptides within the MAP cell wall.…”

Prevalence of Mycobacterium avium Subsp. paratuberculosis in Feral Pigeons (Columba livia) Associated with Difficulties Controlling Paratuberculosis in a Bovine Herd (Fighting Bull Breed)

“…We also found some animals with substantial IT reaction and low antibody values according to OD, with loss of sensitivity to the ELISA test, which could have been because they were in the early stages of the disease [ 7 ]. Nevertheless, sensitivity deficiencies to the ELISA test related to the antigens used in the tests have also been observed, in addition to the variation of sensitivity according to the country when the animals were infected by different strains [ 28 ].…”