Screening of highly expressed mycobacterial genes identifies Rv3615c as a useful differential diagnostic antigen for the Mycobacterium tuberculosis complex.
Tuberculous infections caused by mycobacteria, especially tuberculosis of humans and cattle, are important both clinically and economically. Human populations can be vaccinated with Mycobacterium bovis bacille Calmette-Guérin (BCG), and control measures for cattle involving vaccination are now being actively considered. However, diagnostic tests based on tuberculin cannot distinguish between genuine infection and vaccination with BCG. Therefore, identification of differential diagnostic antigens capable of making this distinction is required, and until now sequence-based approaches have been predominant. Here we explored the link between antigenicity and mRNA expression level, as well as the possibility that we may be able to detect differential antigens by analyzing quantified global transcriptional profiles. We generated a list of 14 candidate antigens that are highly expressed in Mycobacterium tuberculosis and M. bovis under a variety of growth conditions. These candidates were screened in M. bovis-infected and naïve cattle for the ability to stimulate a gamma interferon (IFN-gamma) response. We identified one antigen, Rv3615c, which stimulated IFN-gamma responses in a significant proportion of M. bovis-infected cattle (11 of 30 cattle [37%] [P < 0.01]) but not in naïve or BCG-vaccinated animals. Importantly, the same antigen stimulated IFN-gamma responses in a significant proportion of infected cattle that did not respond to the well-characterized mycobacterial antigens ESAT-6 and CFP-10. Therefore, use of the Rv3615c epitope in combination with previously described differential tests based on ESAT-6 and CFP-10 has the potential to significantly increase diagnostic sensitivity without reducing specificity in BCG-vaccinated populations.
Department of Pathology & Infectious Disease, The Royal Veterinary College, London, United Kingdom.
Ben
B
Sidders
Chris
C
Pirson
Philip J
PJ
Hogarth
R Glyn
RG
Hewinson
Neil G
NG
Stoker
H Martin
HM
Vordermeier
Katie
K
Ewer
D000818:Animals; D000942:Antigens, Bacterial / Q000096:biosynthesis / Q000276:immunology*; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000276:immunology*; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis* / Q000382:microbiology; D002478:Cells, Cultured; D003937:Diagnosis, Differential; D020869:Gene Expression Profiling; D007371:Interferon-gamma / Q000096:biosynthesis; D007963:Leukocytes, Mononuclear / Q000276:immunology; D009163:Mycobacterium bovis / Q000276:immunology* / Q000302:isolation & purification; D009169:Mycobacterium tuberculosis / Q000276:immunology* / Q000302:isolation & purification; D014376:Tuberculosis / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D007371:Interferon-gamma