Production and characterization of anti-DNA-RNA monoclonal antibodies and their application in Listeria detection

Two different heteropolymers cDNA-mRNA and phi X174 DNA-RNA heteroduplexes used as antigens helped obtain murine monoclonal antibodies (MAbs) specific for DNA-RNA hybrids, both having similar immunogenicities. A biotinylated rRNA gene probe specific for the genus Listeria was hybridized with rRNA in teh solution phase and the hybrids formed were captured is microtiter plate wells precoated with the purified zoD3 MAb. The probe-target hybrids were traced with a streptavidin-alkaline phosphatase conjugate. This assay was specific for the genus Listeria and highly sensitive, allowing the detection of as little as 2.5 pg of target rRNA.