Detection of Salmonella species by polymerase chain reaction = Polymerase chain reaction에 의한 Salmonella 속균의 검출

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Title
Detection of Salmonella species by polymerase chain reaction = Polymerase chain reaction에 의한 Salmonella 속균의 검출
Author(s)
Doo Hee Park; Won Yong Kim; Chul Joong Kim; Jum Sool Mah
Bibliographic Citation
Korean Journal of Veterinary Research, vol. 34, no. 1, pp. 115-125
Publication Year
1994
Abstract
A rapid and sensitive method for detection of Salmonella typhi would help in preventing the spread of outbreaks and in clinical diagnosis. In order to develop unique PCR primers to detect Salm. typhi, ribosomal RNA genes from Salm. typhi (Rawlings) were cloned in pUC18. The resulting clone was confirmed by sequencing. The cloned DNA fragment contained the 5S, part of the 23S rRNA genes and the 5S?23S spacer region (EMBL/GenBank accession No. U04734). It was expected that the 5S?23S spacer region is divergent unlike the highly conserved 23S+5S genes. This was confirmed by comparison with the rRNA gene sequences in the EMBL/GenBank database. A pair of PCR primers specific for Salm. typhi was obtained, based on this spacer region sequence. The specificity of this pair of primers was tested with 54 Salm. typhi strains (of 27 different phage types). All these Salm. typhi strains showed the positive 300 bp PCR product with this pair of primers. Six other Salmonella species as well as six other non?Salmonella bacteria were tested and none showed the 300 bp PCR product. The sensitivity of the detection level was 0·1 pg of pure Salm. typhi genomic DNA, or approximately 40 Salm. typhi cells in a spiked food sample. This pair of primers therefore has the potential for development into a diagnostic tool for the rapid diagnosis of typhoid fever.
Keyword
polymerase chain reactionSalmonellaspecific primerPhoESouthern blotDNA probe
ISSN
1225-0198
Publisher
Korea Soc-Assoc-Inst
DOI
http://dx.doi.org/10.1111/j.1365-2672.1996.tb03216.x
Type
Article
Appears in Collections:
1. Journal Articles > Journal Articles
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