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Abstract |
Nonradioactive Hybridization in Microwells Using Chemiluminescence Enzyme Linked Immunosorbent Assay for Detection of RT-PCR-amplified CK19- and CEA-mRNA |
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Cytokeratin 19 (CK19) mRNA and carcinoembryonic antigen (CEA) mRNA as markers for the detection of disseminated carcinoma cells in the bone marrow (BM) and peripheral blood (PB) by reverse transcriptasepolymerase chain reaction (RT-PCR) have been demonstrated in a number of clinical studies on primary breast, gastric, colorectal, lung, and prostate cancer. In this study, we developed chemiluminescence enzyme-linked immunosorbent assays (ELISA) for the detection of RT-PCR-amplified CK19 nRNA and CEA mRNA. The nonradioactive hybridization was performed in a streptavidin-coated microwell with digoxigenin labelled PCR products and a biotin labelled capture probe. Our results showed that chemiluminescence ELISA can be expedient, facile, sensitive and safe for identifing CK19- and CEA-RT-PCR products. These results were proven by sequence. |
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