Use of an arbitrarily primed PCR product in the development of a Campylobacter jejuni-specific PCR

William A. Day, Ian L Pepper, Lynn A. Joens

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Development of a PCR assay for Campylobacter jejuni is based on the isolation of species-specific DNA. An arbitrarily primed PCR incorporating 10-mer primers was used to generate fingerprints of C. jejuni M129 genomic DNA. Fingerprint products were then screened individually for their species specificity in dot blot hybridizations with 6 C. jejuni isolates, 4 Campylobacter species other than C. jejuni, and 27 enteric bacterial species other than Campylobacter spp. A 486-bp fingerprint product hybridized specifically to C. jejuni DNA under stringent conditions; no binding to Campylobacter DNA other than that of C. jejuni or to DNA from enteric bacteria was detected. The 486-bp fingerprint product was sequenced, and primers corresponding to three overlapping regions of the DNA probe were synthesized. Evaluation of the three primer pairs for specificity to C. jejuni DNA identified an oligonucleotide primer pair which amplified a 265- bp product from six C. jejuni isolates only. In sensitivity studies using a crude M129 lysate as the template, the C. jejuni-specific PCR amplified the 265-bp product in a lysate with as few as 100 bacteria.

Original languageEnglish (US)
Pages (from-to)1019-1023
Number of pages5
JournalApplied and Environmental Microbiology
Volume63
Issue number3
StatePublished - Mar 1997

Fingerprint

Campylobacter jejuni
product development
DNA
Polymerase Chain Reaction
Dermatoglyphics
Campylobacter
bacterium
product
Species Specificity
DNA Primers
DNA probes
DNA Probes
DNA primers
genomics
Enterobacteriaceae
nucleic acid hybridization
intestinal microorganisms
probe
assay
Bacteria

ASJC Scopus subject areas

  • Environmental Science(all)
  • Biotechnology
  • Microbiology

Cite this

Use of an arbitrarily primed PCR product in the development of a Campylobacter jejuni-specific PCR. / Day, William A.; Pepper, Ian L; Joens, Lynn A.

In: Applied and Environmental Microbiology, Vol. 63, No. 3, 03.1997, p. 1019-1023.

Research output: Contribution to journalArticle

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