Article Abstract:
A chemiluminescent or a colorimetric substrate for the nonisotopic detection of the ligase chain reaction (LCR) was used the polymerase chain reaction (PCR)-coupled LCR indicated a sensitivity of 10 CFU of Listeria monocytogenes. The detection technique with the chemiluminescent substrate Lumi-Phos 530 allowed detection of the LCR products in less than three hours so as to complete it within 10 hours. An there was a gradual increase in the number of LCR cycles (LCR > 10) that were examined, a ligated product was noticed with the nonisotopic detection mode when L. inocera was a target, and high background signals arose after exposure of the Lumi-Phos 530 reaction to polaroid type 612 film for 10 minutes or longer. An exposure time of 5 minutes was ideal for optimum results.
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Article Abstract:
Two pairs of primers were designed for use in a ligase chain reaction (LCR) assay to discriminate Listeria monocytogenes from other Listeria species. The primers were based on the 16s rRNA gene containing a single base-pair difference in theV9 region. Prior to LCR, the probes were amplified using the polymerase chain reaction. Tests on 19 different Listeria species showed that the procedure could effectively discriminate between species, and can therefore be a useful diagnostic tool for detecting Listeria monocytogenes.
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Article Abstract:
The discrimination of Listeria monocytogenes from Listeria innocua by differences in the 16S rRna genes is discussed. The genes were amplified by polymerase chain reaction and cloned from the bacilli and their random amplified polymorphic DNA (RAPD) patterns were established. The ability of RAPD for resolving the origin of the Listeria isolates was satisfactorily demonstrated.
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