PCE-ELISA در تشخيص عفونت خوني ناشي از باكتريهاي گرم منفي

Development of a PCR-ELISA for Diagnosing Blood Infection Caused by Gram Negative Bacteria

A polymerase chain reaction enzyme-linked immunosorbent assay (PCR-ELISA) was used to measure levels of circulating gram negative bacterial DNA in serum. The PCR reaction was based on primers from the 16S rRNA gene. The product was digoxigenylated during PCR and this was the tag to which antibody was bound in the subsequent ELISA. The product was bound to a streptavidin-coated microtitration plate which was mediated by a biotinylated capture probe. The optical density (OD) endpoint was <0.1 in 10 sera from healthy people and also it was <0.1 in 45 out of 50 patients with blood infection other than gram negative bacteria (specificity = 90.9%). In 43 cases of proven gram negative bacteremia an OD >0.1 was achieved in 40 patients (sensitivity = 93%). This assay validated the concept of diagnosing gram negative bacteria ; by measuring levels of circulating DNA in serum.