Degradation of cardiac troponin I in serum complicates comparisons of cardiac troponin I assays.
BACKGROUND
Up to a 20-fold variation in serum cardiac troponin I (cTnI) concentration may be observed for a given patient sample with different analytical methods. Because more limited variation is seen for control materials and for purified cTnI, we explored the possibility that cTnI was present in altered forms in serum.


METHODS
We used four recombinantly engineered cTnI fragments to study the regions of cTnI recognized by the Stratus(R), Opus(R), and ACCESS(R) immunoassays. The stability of these regions in serum was analyzed with Western blot.


RESULTS
The measurement of several control materials and different forms of purified cTnI using selected commercial assays demonstrated five- to ninefold variation. Both the Stratus and Opus assays recognized the N-terminal portion (NTP) of cTnI, whereas the ACCESS assay recognized the C-terminal portion (CTP) of cTnI. Incubation of recombinant cTnI in normal human serum produced a marked decrease in cTnI concentration as determined with the ACCESS, but not the Stratus, immunoassay. Western blot analysis of the same samples using cTnI NTP- and CTP-specific antibodies demonstrated preferential degradation of the CTP of cTnI.


CONCLUSIONS
The availability of serum cTnI epitopes is markedly affected by the extent of ligand degradation. The N-terminal half of the cTnI molecule was found to be the most stable region in human serum. Differential degradation of cTnI is a key factor in assay-to-assay variation.
Shi
Q
Q
Spectral Diagnostics Inc., 135-2 The West Mall, Toronto, Ontario, Canada M9C 1C2. qshi@ica.net
Ling
M
M
Zhang
X
X
Zhang
M
M
Kadijevic
L
L
Liu
S
S
Laurino
J P
JP
D015153:Blotting, Western; D006801:Humans; D007118:Immunoassay; D009206:Myocardium / Q000378:metabolism*; D011933:Reagent Kits, Diagnostic; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000097:blood; D019210:Troponin I / Q000096:biosynthesis / Q000097:blood*
D011933:Reagent Kits, Diagnostic; D011994:Recombinant Proteins; D019210:Troponin I