Qualitative bedside assay of increased human serum myoglobin by sandwich dot-immunogold filtration for the diagnosis of acute myocardial infarction.
We isolated and purified myoglobin (MYO) from human fresh skeletal muscle and prepared monoclonal and polyclonal antibody from it. A sandwich dot-immunogold filtration assay (DIGFA) for the detection of MYO was developed by using affinity purified sheep anti-MYO antibody as the first antibody for coating nitrocellulose membranes (NCMs; the support) and colloidal gold labelled monoclonal antibody (H3) as the second antibody (an indicator). The test can be completed in 3 min without incubation or any equipment. A reddish dot, indicating positivity, is obvious to the naked eye. No interferences from bilirubin, hemoglobin, rheumatoid factors and lipid were found. In order to use undiluted serum, the detection limit was set at 100 microg of MYO/l. Concentrations up to 30,000 microg/l can be measured without getting a "hook" effect. Serum MYO levels in 53 patients with acute myocardial infarction (AMI), 100 healthy individuals, seven patients with chest pain but without myocardial ischemia and in 39 patients with renal insufficiency were measured simultaneously by DIGFA and enzyme-linked immunosorbent assay (ELISA). All serum samples from patients had MYO concentrations above 100 microg/l by ELISA and were positive by DIGFA. Serum creatinine values were related to MYO test results. Healthy individuals had MYO levels below 85 microg/l by ELISA and were negative by DIGFA.
Qiaojia
H
H
Center for Medical Laboratory Science, Fuzhou General (Dong Fang) Hospital, Fujian, China. ammnfghp@med1.bmi.ac.cn
Yuchai
T
T
Weiping
L
L
Shuxuan
Y
Y
Xiaopeng
L
L
Bing
X
X
Yushui
W
W
Li
L
L
Zhongyong
Z
Z
D000328:Adult; D000368:Aged; D000911:Antibodies, Monoclonal; D003402:Creatine Kinase / Q000097:blood; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005374:Filtration; D006801:Humans; D007150:Immunohistochemistry; D007527:Isoenzymes; D008297:Male; D008875:Middle Aged; D009203:Myocardial Infarction / Q000175:diagnosis*; D009211:Myoglobin / Q000097:blood* / Q000276:immunology; D019095:Point-of-Care Systems
D000911:Antibodies, Monoclonal; D007527:Isoenzymes; D009211:Myoglobin; D003402:Creatine Kinase