Detection of Salmonella typhi in the blood of patients with typhoid fever by polymerase chain reaction

J H Song, H Cho, M Y Park, D S Na, H B Moon, C H Pai

Research output: Contribution to journalArticlepeer-review

139 Scopus citations

Abstract

A polymerase chain reaction (PCR)-based test was developed for the detection of Salmonella typhi in the blood specimens from patients with typhoid fever. Two pairs of oligonucleotide primers were designed to amplify a 343-bp fragment of the flagellin gene of S. typhi. Amplified products were analyzed by agarose gel electrophoresis and Southern blot hybridization by using a 32P-labeled 40-base probe internal to the amplified DNA. The nested PCR with two pairs of primers could detect 10 organisms of S. typhi as determined by serial dilutions of DNA from S. typhi. The peripheral mononuclear cells from 11 of 12 patients with typhoid fever confirmed by blood culture were positive for DNA fragment of the flagellin gene of S. typhi, whereas 10 blood specimens of patients with other febrile diseases were negative. With the nested PCR, S. typhi DNAs were detected from blood specimens of four patients with suspected typhoid fever on the basis of clinical features but with negative cultures. We suggest that the PCR technique could be used as a novel diagnostic method of typhoid fever, particularly in culture-negative cases.

Original languageEnglish (US)
Pages (from-to)1439-43
Number of pages5
JournalJournal of clinical microbiology
Volume31
Issue number6
DOIs
StatePublished - Jun 1993

Keywords

  • Bacteremia/diagnosis
  • Base Sequence
  • DNA Probes
  • DNA, Bacterial/genetics
  • Flagellin/genetics
  • Genes, Bacterial
  • Humans
  • Molecular Sequence Data
  • Polymerase Chain Reaction/methods
  • Salmonella typhi/genetics
  • Sensitivity and Specificity
  • Typhoid Fever/diagnosis

PubMed: MeSH publication types

  • Journal Article
  • Research Support, Non-U.S. Gov't

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