Immunological capture of nucleic acid hybrids and application to nonradioactive DNA probe assay.
Antibodies specific for DNA:RNA hybrids were coated onto polystyrene test tubes and applied to hybridization assays involving DNA and RNA. Synthetic DNA probes complementary to 16S rRNA of Campylobacter were labeled with biotin and hybridized to ribosomal RNA directly in lysates of bacterial cells. After hybridization, DNA:RNA hybrids were captured with immobilized anti-DNA:RNA antibody, and the biotinylated probe was detected with streptavidin-horseradish peroxidase (EC 1.11.1.7) conjugate. The assay was optimized to detect as few as 70,000 Campylobacter cells in a sample. We compared the utility of this hybridization assay with that of conventional microbiology methods by examination of 1448 stool samples from hospital clinical laboratories. The DNA hybridization assay had a sensitivity of 98.7% (75/76) and a specificity of 98.2% (1347/1372) and overall agreed with 98.2% of the conventional results for a test population that had a 5.2% incidence (76/1448) of Campylobacter infection. The assay is simple to perform and yields results within 2.5 h.
D000906:Antibodies; D001424:Bacterial Infections / Q000382:microbiology*; D002168:Campylobacter fetus / Q000235:genetics; D003429:Cross Reactions; D003967:Diarrhea / Q000382:microbiology*; D005243:Feces / Q000382:microbiology; D006801:Humans; D008954:Models, Biological; D009693:Nucleic Acid Hybridization; D012329:RNA, Bacterial / Q000032:analysis*; D012335:RNA, Ribosomal / Q000032:analysis*; D013053:Spectrophotometry / Q000379:methods
D000906:Antibodies; D012329:RNA, Bacterial; D012335:RNA, Ribosomal