Development of a sensitive enzyme-linked immunosorbent assay for measurement of DNase I in human serum.
BACKGROUND
Serum deoxyribonuclease I (DNase I) activity was reported to increase in the early phase after onset of acute myocardial infarction (AMI). Up to now, DNase I activity has been quantified by the single radial enzyme diffusion (SRED) method, which unfortunately requires a long incubation time. Therefore it is necessary to develop another assay suitable for measurement of serum DNase I concentrations in a clinical setting.


METHODS
A sandwich ELISA was established for measurement of DNase I protein using a polyclonal antibody directed against DNase I protein and a biotinylated monoclonal for subsequent detection. Concentrations of serum DNase I protein were measured in healthy individuals and patients with AMI.


RESULTS
This method was as precise as SRED, and took less time than SRED. A significant correlation was observed between DNase I concentration and enzyme activity (r=0.839; P<0.001). The average of serum DNase I in AMI patients within 0-12 h of chest pain was significantly higher than that in healthy individuals (P<0.001), and decreased with time.


CONCLUSIONS
We have developed a sensitive ELISA capable of measuring DNase I protein concentrations. This method may be a useful alternative to SRED as an aid to diagnosis of AMI based on the serum DNase I level.
Nakajima
Tamiko
T
Department of Legal Medicine, Gunma University, Graduate School of Medicine, Gunma, Japan.
Takagi
Rie
R
Tajima
Yutaka
Y
Makita
Chikako
C
Kominato
Yoshihiko
Y
Kuribara
Jun
J
Ohshima
Shigeru
S
Tada
Hiroshi
H
Tsurugaya
Hideki
H
Kobayashi
Yasuyuki
Y
Takeshita
Haruo
H
Kawai
Yasuyuki
Y
Yasuda
Toshihiro
T
D000328:Adult; D000906:Antibodies / Q000276:immunology; D000918:Antibody Specificity; D003850:Deoxyribonuclease I / Q000097:blood* / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D009203:Myocardial Infarction / Q000097:blood / Q000201:enzymology; D012680:Sensitivity and Specificity; D013997:Time Factors
D000906:Antibodies; D003850:Deoxyribonuclease I