Partial inhibition of amplifications by primers of EHEC genes

I. Gadó, Zs Ruzsics, I. Tóth, Margit Király, Hedda Milch

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Diagnostic value of multiplex polymerase chain reaction (PCR) was examined by using three primer pairs, specific for the common conserved region of stx1 and stx2, eae and an enterohaemolysin A gene (ehxA). The sensitivity in respect of each amplicon decreased with three exponents comparing to the individual PCR reactions. These PCR reactions were partially inhibited by the presence of certain additional primers. This inhibitory effect was template-concentration dependent, and was partially balanced by usage of increased amount of dNTP. Taq DNA polymerase in a range of 0.3-1.25 U/reaction did not influence the inhibition. The same inhibiton was detected if the annealing temperature was changed from 48°C to 57°C. Pairs of EHEC primers inhibited a Salmonella enteritidis virulence-plasmid specific gene amplification, as well. Theoretical inhibiting effects were predicted by Primer Premier software but our observations can be sufficiently explained neither by the competitions between the specific and aspecific amplifications nor by the inhibition caused by dimerization of primers.

Original languageEnglish
Pages (from-to)239-252
Number of pages14
JournalActa Microbiologica et Immunologica Hungarica
Volume45
Issue number2
Publication statusPublished - 1998

ASJC Scopus subject areas

  • Immunology and Microbiology(all)
  • Medicine(all)
  • Microbiology

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