Novel recombinant RD2- and RD11-encoded Mycobacterium tuberculosis antigens are potential candidates for diagnosis of tuberculosis infections in BCG-vaccinated individuals.
Proteins encoded by region of deletions (RD) of Mycobacterium tuberculosis are useful in development of vaccines and diagnostic reagents. In the present study, six M. tuberculosis genes from RD2 and RD11, rv1978, nrdf1, mpt64, cfp-21, ppe57 and ppe59, were cloned and overexpressed in Escherichia coli. All six purified recombinant proteins could distinguish tuberculosis (TB) patients and latent TB infected subjects (LTBI), or called subclinical TB infection, from BCG-vaccinated healthy controls by T-cell IFN-gamma releasing ELISPOT. ELISPOT of Rv1978, NrdF1, Mpt64, CFP-21, Ppe57 and Ppe59 achieved sensitivities of 59%, 60%, 82%, 48%, 59% and 47% respectively in the detection of active TB and specificities of 94%, 90%, 76%, 93%, 100% and 93% respectively in BCG-vaccinated healthy controls. Combination of Ppe57 or NrdF1 with early secreted antigen target 6 (ESAT-6) or 10-kDa culture filtrate protein (CFP-10) in the IFN-gamma releasing ESLIPOT assay could increase the sensitivities in detecting active TB, for ESAT-6 from 82.1% to 85.7% or 92.9% (P=0.5 or 0.03, respectively) and for CFP-10 from 67.9% to 78.6% or 83.9%, respectively (both P<0.05). The high sensitivities, specificities and promising antigenic combination of NrdF1 and Ppe57 in detection of TB in BCG-vaccinated controls suggest their potential application in TB diagnosis.
Chen
Jiazhen
J
State Key Laboratory of Genetic Engineering, Institute of Genetics, Fudan University, Shanghai 200433, China.
Su
Xiaodi
X
Zhang
Ying
Y
Wang
Sen
S
Shao
Linyun
L
Wu
Jing
J
Wang
Feifei
F
Zhang
Shu
S
Wang
Jiuling
J
Weng
Xinhua
X
Wang
Honghai
H
Zhang
Wenhong
W
D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000942:Antigens, Bacterial / Q000235:genetics; D001500:BCG Vaccine / Q000276:immunology*; D002648:Child; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D015870:Gene Expression; D006801:Humans; D007124:Immunoenzyme Techniques / Q000379:methods*; D007371:Interferon-gamma / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D011994:Recombinant Proteins / Q000235:genetics; D012680:Sensitivity and Specificity; D013601:T-Lymphocytes / Q000276:immunology; D014376:Tuberculosis / Q000175:diagnosis*; D055815:Young Adult
D000942:Antigens, Bacterial; D001500:BCG Vaccine; D011994:Recombinant Proteins; C072681:Mycobacterium tuberculosis antigens; D007371:Interferon-gamma