A multiplexed and miniaturized serological tuberculosis assay identifies antigens that discriminate maximally between TB and non-TB sera.
We have developed a multiplexed and miniaturized TB serological assay with the aim of identifying (combinations of) antigens that maximally discriminate between TB and non-TB patients. It features a microarray accommodating 54 TB antigens, less than 1 microl serum consumption and an indirect immunofluorescence detection protocol. With a panel of 20 TB and 80 non-TB sera we ranked combinations of TB antigens with respect to sensitivity and specificity of TB detection by means of logistic step-forward regression analysis. The highest-ranking TB antigen combination had an area-under-the-curve of the receiver-operator-characteristics (ROC) of 0.95. We also identified an antigen that on its own provided good specificity and sensitivity of TB detection (Ara6-BSA; area-under-the-ROC curve: 0.90). These area-under-the-ROC curve values are exceptionally high for a serological TB assay. We conclude that TB antigen microarrays permit rapid identification of TB antigens that, either alone or in combination, discriminate maximally between TB and non-TB patients and that such identification provides an excellent starting point for developing point-of-care diagnostic assays.
Tong
Miao
M
Department of Molecular Cell Biology, Leiden University Medical Center, Leiden, The Netherlands.
Jacobi
Catharina E
CE
van de Rijke
Frans M
FM
Kuijper
Sjoukje
S
van de Werken
Sjaak
S
Lowary
Todd L
TL
Hokke
Cornelis H
CH
Appelmelk
Ben J
BJ
Nagelkerke
Nico J D
NJ
Tanke
Hans J
HJ
van Gijlswijk
Rob P M
RP
Veuskens
Jacques
J
Kolk
Arend H J
AH
Raap
Anton K
AK
D000907:Antibodies, Bacterial / Q000276:immunology; D000918:Antibody Specificity; D000942:Antigens, Bacterial / Q000276:immunology*; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D006801:Humans; D007106:Immune Sera / Q000276:immunology*; D008969:Molecular Sequence Data; D009169:Mycobacterium tuberculosis / Q000276:immunology*; D009844:Oligosaccharides / Q000737:chemistry / Q000378:metabolism; D040081:Protein Array Analysis; D012372:ROC Curve; D012680:Sensitivity and Specificity; D012698:Serologic Tests / Q000379:methods*; D014376:Tuberculosis / Q000097:blood* / Q000175:diagnosis / Q000276:immunology* / Q000382:microbiology
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D007106:Immune Sera; D009844:Oligosaccharides