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. 1993 Jun;31(6):1439–1443. doi: 10.1128/jcm.31.6.1439-1443.1993

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

J H Song 1, H Cho 1, M Y Park 1, D S Na 1, H B Moon 1, C H Pai 1
PMCID: PMC265558  PMID: 8314983

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.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Edelman R., Levine M. M. Summary of an international workshop on typhoid fever. Rev Infect Dis. 1986 May-Jun;8(3):329–349. doi: 10.1093/clinids/8.3.329. [DOI] [PubMed] [Google Scholar]
  2. Frankel G., Newton S. M., Schoolnik G. K., Stocker B. A. Intragenic recombination in a flagellin gene: characterization of the H1-j gene of Salmonella typhi. EMBO J. 1989 Oct;8(10):3149–3152. doi: 10.1002/j.1460-2075.1989.tb08468.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Frankel G., Newton S. M., Schoolnik G. K., Stocker B. A. Unique sequences in region VI of the flagellin gene of Salmonella typhi. Mol Microbiol. 1989 Oct;3(10):1379–1383. doi: 10.1111/j.1365-2958.1989.tb00119.x. [DOI] [PubMed] [Google Scholar]
  4. Gilman R. H., Terminel M., Levine M. M., Hernandez-Mendoza P., Hornick R. B. Relative efficacy of blood, urine, rectal swab, bone-marrow, and rose-spot cultures for recovery of Salmonella typhi in typhoid fever. Lancet. 1975 May 31;1(7918):1211–1213. doi: 10.1016/s0140-6736(75)92194-7. [DOI] [PubMed] [Google Scholar]
  5. Guerra-Caceres J. G., Gotuzzo-Herencia E., Crosby-Dagnino E., Miro-Quesada M., Carrillo-Parodi C. Diagnostic value of bone marrow culture in typhoid fever. Trans R Soc Trop Med Hyg. 1979;73(6):680–683. doi: 10.1016/0035-9203(79)90020-8. [DOI] [PubMed] [Google Scholar]
  6. Hoffman S. L., Edman D. C., Punjabi N. H., Lesmana M., Cholid A., Sundah S., Harahap J. Bone marrow aspirate culture superior to streptokinase clot culture and 8 ml 1:10 blood-to-broth ratio blood culture for diagnosis of typhoid fever. Am J Trop Med Hyg. 1986 Jul;35(4):836–839. doi: 10.4269/ajtmh.1986.35.836. [DOI] [PubMed] [Google Scholar]
  7. Levine M. M., Grados O., Gilman R. H., Woodward W. E., Solis-Plaza R., Waldman W. Diagnostic value of the Widal test in areas endemic for typhoid fever. Am J Trop Med Hyg. 1978 Jul;27(4):795–800. doi: 10.4269/ajtmh.1978.27.795. [DOI] [PubMed] [Google Scholar]
  8. Rubin F. A., Kopecko D. J., Noon K. F., Baron L. S. Development of a DNA probe to detect Salmonella typhi. J Clin Microbiol. 1985 Oct;22(4):600–605. doi: 10.1128/jcm.22.4.600-605.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Rubin F. A., Kopecko D. J., Sack R. B., Sudarmono P., Yi A., Maurta D., Meza R., Moechtar M. A., Edman D. C., Hoffman S. L. Evaluation of a DNA probe for identifying Salmonella typhi in Peruvian and Indonesian bacterial isolates. J Infect Dis. 1988 May;157(5):1051–1053. doi: 10.1093/infdis/157.5.1051. [DOI] [PubMed] [Google Scholar]
  10. Rubin F. A., McWhirter P. D., Burr D., Punjabi N. H., Lane E., Kumala S., Sudarmono P., Pulungsih S. P., Lesmana M., Tjaniadi P. Rapid diagnosis of typhoid fever through identification of Salmonella typhi within 18 hours of specimen acquisition by culture of the mononuclear cell-platelet fraction of blood. J Clin Microbiol. 1990 Apr;28(4):825–827. doi: 10.1128/jcm.28.4.825-827.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Rubin F. A., McWhirter P. D., Punjabi N. H., Lane E., Sudarmono P., Pulungsih S. P., Lesmana M., Kumala S., Kopecko D. J., Hoffman S. L. Use of a DNA probe to detect Salmonella typhi in the blood of patients with typhoid fever. J Clin Microbiol. 1989 May;27(5):1112–1114. doi: 10.1128/jcm.27.5.1112-1114.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Wei L. N., Joys T. M. Covalent structure of three phase-1 flagellar filament proteins of Salmonella. J Mol Biol. 1985 Dec 20;186(4):791–803. doi: 10.1016/0022-2836(85)90397-3. [DOI] [PubMed] [Google Scholar]
  13. Wicks A. C., Cruickshank J. G., Musewe N. Observations on the diagnosis of typhoid fever in an endemic area. S Afr Med J. 1974 Jul 6;48(32):1368–1370. [PubMed] [Google Scholar]

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