PCR detection of a putative N-acetylmuramidase gene from Listeria ivanovii facilitates its rapid identification.
Listeria ivanovii is a Gram-positive bacterial pathogen that is capable of causing abortions and stillbirths in farm animals, particularly sheep and cattle. In terms of morphological, biochemical and molecular characteristics, L. ivanovii resembles other Listeria species such as L. monocytogenes, a pathogen of both man and animals. In this study, through comparative analysis of genomic DNA from the six Listeria species, a L. ivanovii specific clone (liv22-228) containing a 946 bp insert was isolated. This clone contained the 5' ends of two divergently transcribed L. ivanovii genes and an intergenic spacer region, similar in organization to homologous regions from the L. innocua and L. monocytogenes genomes. Regions of low homology in the clone were identified by comparing to the L. innocua and L. monocytogenes genomes, and oligonucleotide primers (liv22-228F and liv22-228R) were designed. These primers amplified a 463 bp band from genomic DNA of L. ivanovii strains only, but not from other Listeria species or common bacteria. Thus, PCR employing L. ivanovii specific primers (liv22-228F and liv22-228R) provides a useful and straightforward method for rapid and precise determination of L. ivanovii.
Liu
Dongyou
D
Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, P.O. Box 6100, Mississippi, MS 39762, USA.
Ainsworth
A Jerald
AJ
Austin
Frank W
FW
Lawrence
Mark L
ML
D000034:Abortion, Veterinary / Q000382:microbiology; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D002418:Cattle Diseases / Q000382:microbiology*; D003001:Cloning, Molecular; D017931:DNA Primers; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D005260:Female; D006026:Glycoside Hydrolases / Q000737:chemistry / Q000235:genetics*; D008087:Listeria / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification; D008088:Listeriosis / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary; D011247:Pregnancy
D017931:DNA Primers; D004269:DNA, Bacterial; D006026:Glycoside Hydrolases; C026255:endo-N-acetylmuramidase