Transcriptional enhancement of the Listeria monocytogenes PCR and simple immunoenzymatic assay of the product using anti-RNA:DNA antibodies.
A method was developed to enhance the sensitivity of a Listeria monocytogenes PCR detection system by in vitro transcription of amplicons incorporating bacteriophage T7 RNA polymerase promoter sequences in one of the priming oligonucleotides. The resulting transcript can be detected by hybridization with a DNA probe immobilized in the wells of a microtiter plate, followed by immunoenzymatic assay of the RNA-DNA hybrids with an anti-RNA-DNA hybrid antibody. This highly sensitive method was reactive in the assay of various L. monocytogenes isolates but not with other Listeria or non-Listeria species.
Blais
B W
BW
Laboratory Services Division, Agriculture and Agri-Food Canada, Ottawa, Ontario.
D000907:Antibodies, Bacterial; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D015342:DNA Probes; D004269:DNA, Bacterial / Q000235:genetics / Q000276:immunology; D005069:Evaluation Studies as Topic; D007124:Immunoenzyme Techniques; D008089:Listeria monocytogenes / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000706:statistics & numerical data; D012329:RNA, Bacterial / Q000235:genetics / Q000276:immunology; D012680:Sensitivity and Specificity; D014158:Transcription, Genetic
D000907:Antibodies, Bacterial; D017931:DNA Primers; D015342:DNA Probes; D004269:DNA, Bacterial; D012329:RNA, Bacterial