Tumor necrosis factor-alpha in serum of patients with inflammatory bowel disease as measured by a highly sensitive immuno-PCR.
BACKGROUND
The significance of serum concentrations of tumor necrosis factor-alpha (TNF-alpha) in the pathogenesis of inflammatory bowel disease (IBD) is uncertain. We measured TNF-alpha in serum from IBD patients by immuno-PCR to analyze the relationship between TNF-alpha and pathophysiologic state in IBD.


METHODS
Serum samples were collected from 54 healthy blood donors, 29 patients with ulcerative colitis (UC; 46 samples), and 7 patients with Crohn disease (CD; 8 samples). DNA label was generated by PCR amplification using biotinylated primer and was bound with streptavidin to biotinylated third antibody. TNF-alpha sandwiched by antibodies was detected by PCR amplification of the DNA label.


RESULTS
TNF-alpha could be measured in all samples. The median serum concentration in IBD patients overall was approximately 390-fold higher than in healthy donors (median increase, 380-fold for UC, 640-fold for CD). The median serum TNF-alpha concentration was 1.7-fold higher in the active stage of UC than in the inactive stage (P <0.05), and this difference could be detected in individual patients.


CONCLUSIONS
Sensitive measurement of serum TNF-alpha could provide an important pathophysiologic marker for the presence and activity of IBD.
Komatsu
M
M
Division of Laboratory Diagnosis, Department of Clinical Laboratory Medicine, Sapporo Medical University, School of Medicine, South-I, West-16, Chuo-ku, Sapporo 060-8543, Japan.
Kobayashi
D
D
Saito
K
K
Furuya
D
D
Yagihashi
A
A
Araake
H
H
Tsuji
N
N
Sakamaki
S
S
Niitsu
Y
Y
Watanabe
N
N
D000328:Adult; D000368:Aged; D000906:Antibodies; D015415:Biomarkers / Q000097:blood; D019921:Biotinylation; D003093:Colitis, Ulcerative / Q000097:blood / Q000473:pathology; D003424:Crohn Disease / Q000097:blood / Q000473:pathology; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D015212:Inflammatory Bowel Diseases / Q000097:blood* / Q000473:pathology; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D019809:Streptavidin; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis*
D000906:Antibodies; D015415:Biomarkers; D014409:Tumor Necrosis Factor-alpha; D019809:Streptavidin