A novel immuno-polymerase chain reaction protocol incorporating a highly purified streptavidin-DNA conjugate.
We have developed an immuno-polymerase chain reaction protocol which includes a highly purified streptavidin-DNA conjugate. The protocol comprises standard ELISA methodology and washing buffers together with a real-time PCR read-out system. The conjugate was employed in both indirect and capture assay formats, which can be completed in a single day using standard laboratory equipment. The immuno-PCR is reproducible, with a larger dynamic range and a sensitivity several orders of magnitude greater than the corresponding conventional ELISA. The minimum concentration of analyte detected was of the order of 1 pg/mL These characteristics should contribute to the adoption of immuno-PCR by research and clinical laboratories.
Shan
Jianguo
J
AGEN Biomedical Limited, Acacia Ridge, Queensland, Australia.
Toye
Philip
P
D000818:Animals; D019921:Biotinylation; D004247:DNA / Q000737:chemistry*; D004184:Dirofilariasis / Q000175:diagnosis; D004283:Dog Diseases / Q000175:diagnosis; D004285:Dogs; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005338:Fibrin Fibrinogen Degradation Products / Q000032:analysis; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D019809:Streptavidin / Q000737:chemistry*
D005338:Fibrin Fibrinogen Degradation Products; C036309:fibrin fragment D; D004247:DNA; D019809:Streptavidin