A 16S rRNA-based DNA probe and PCR method specific forListeria ivanovii

Abstract

A 16S rRNA-based DNA probe and polymerase chain reaction (PCR) method was developed for identification and rapid detection of Listeria ivanovii. The probe (R-1) is 5'-GTAGTGACGCATGTCATCAC-3' corresponding to positions 185-204 in the L. ivanovii 16S rRNA sequence. DNA hybridization results indicated that R-1 probe only reacted with L. ivanovii, and not with six other species of Listeria or other bacteria tested. The PCR method using R-1 and a reverse primer, R-2, was positive with all eight strains of L. ivanovii tested but was negative with six other species of Listeria, including nine strains of L. monocytogenes, and 20 other taxonomically related bacteria tested. In our PCR method, starting with whole bacterial cells, only 3 h were required for the PCR assay and 1 h for electrophoresis without any additional time for DNA isolation and DNA hybridization. This PCR method detected as few as 4 cells of L. ivanovii in pure cultures and 4-40 cells of L. ivanovii in inoculated and diluted mouse feed, blood, or faeces samples.