Skip to main content
PMC home page
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1985 Oct;22(4):600–605. doi: 10.1128/jcm.22.4.600-605.1985

Development of a DNA probe to detect Salmonella typhi.

F A Rubin, D J Kopecko, K F Noon, L S Baron
PMCID: PMC268475  PMID: 3001134

Abstract

This study was undertaken to identify a DNA sequence that could be used to facilitate the diagnostic identification of Salmonella typhi, the causative agent of typhoid fever. All virulent S. typhi strains encode a relatively unique capsular antigen termed the virulence (Vi) antigen. Two distinct genetic loci, viaA and viaB, are involved in the synthesis of this antigen. The structural genes, located at viaB, were considered as a possible specific DNA probe. The viaB locus, contained in a recombinant cosmid, was subcloned to various plasmid vectors for this purpose. Selected viaB-region DNA fragments were then analyzed for specificity in DNA colony hybridization reactions with more than 170 strains representing a variety of enteric bacteria. An 8.6-kilobase EcoRI fragment was highly specific for the viaB gene region and was considered a good hybridization probe. This DNA probe should prove useful in rapid diagnostic assays set up to detect S. typhi in mixed bacterial samples (e.g., stools) within a few hours of specimen collection.

Full text

PDF
601

Images in this article

603Image
on p.603

Selected References

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

  1. Baron L. S., Kopecko D. J., McCowen S. M., Snellings N. J., Johnson E. M., Reid W. C., Life C. A. Genetic and molecular studies of the regulation of atypical citrate utilization and variable Vi antigen expression in enteric bacteria. Basic Life Sci. 1982;19:175–194. doi: 10.1007/978-1-4684-4142-0_16. [DOI] [PubMed] [Google Scholar]
  2. Boileau C. R., d'Hauteville H. M., Sansonetti P. J. DNA hybridization technique to detect Shigella species and enteroinvasive escherichia coli. J Clin Microbiol. 1984 Nov;20(5):959–961. doi: 10.1128/jcm.20.5.959-961.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bolivar F. Construction and characterization of new cloning vehicles. III. Derivatives of plasmid pBR322 carrying unique Eco RI sites for selection of Eco RI generated recombinant DNA molecules. Gene. 1978 Oct;4(2):121–136. doi: 10.1016/0378-1119(78)90025-2. [DOI] [PubMed] [Google Scholar]
  4. Ditta G., Stanfield S., Corbin D., Helinski D. R. Broad host range DNA cloning system for gram-negative bacteria: construction of a gene bank of Rhizobium meliloti. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7347–7351. doi: 10.1073/pnas.77.12.7347. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Echarti C., Hirschel B., Boulnois G. J., Varley J. M., Waldvogel F., Timmis K. N. Cloning and analysis of the K1 capsule biosynthesis genes of Escherichia coli: lack of homology with Neisseria meningitidis group B DNA sequences. Infect Immun. 1983 Jul;41(1):54–60. doi: 10.1128/iai.41.1.54-60.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fitts R., Diamond M., Hamilton C., Neri M. DNA-DNA hybridization assay for detection of Salmonella spp. in foods. Appl Environ Microbiol. 1983 Nov;46(5):1146–1151. doi: 10.1128/aem.46.5.1146-1151.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hill W. E., Payne W. L., Aulisio C. C. Detection and enumeration of virulent Yersinia enterocolitica in food by DNA colony hybridization. Appl Environ Microbiol. 1983 Sep;46(3):636–641. doi: 10.1128/aem.46.3.636-641.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hill W. E., Payne W. L., Crouch R. J., Davis V. M., English L. L., Ferreira J. L., Gemski P., Jagow J. A., Moseley S. L., Noah C. W. Genetic methods for the detection of microbial pathogens. Identification of enterotoxigenic Escherichia coli by DNA colony hybridization: collaborative study. J Assoc Off Anal Chem. 1984 Jul-Aug;67(4):801–807. [PubMed] [Google Scholar]
  9. Hohn B., Collins J. A small cosmid for efficient cloning of large DNA fragments. Gene. 1980 Nov;11(3-4):291–298. doi: 10.1016/0378-1119(80)90069-4. [DOI] [PubMed] [Google Scholar]
  10. JOHNSON E. M., KRAUSKOPF B., BARON L. S. GENETIC MAPPING OF VI AND SOMATIC ANTIGENIC DETERMINANTS IN SALMONELLA. J Bacteriol. 1965 Aug;90:302–308. doi: 10.1128/jb.90.2.302-308.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Johnson E. M., Krauskopf B., Baron L. S. Genetic analysis of the ViA-his chromosomal region in Salmonella. J Bacteriol. 1966 Nov;92(5):1457–1463. doi: 10.1128/jb.92.5.1457-1463.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. KRISHNAPILLAI V., BARON L. S. ALTERATIONS IN THE MOUSE VIRULENCE OF SALMONELLA TYPHIMURIUM BY GENETIC RECOMBINATION. J Bacteriol. 1964 Mar;87:598–605. doi: 10.1128/jb.87.3.598-605.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kaper J. B., Campen R. K., Seidler R. J., Baldini M. M., Falkow S. Cloning of the thermostable direct or Kanagawa phenomenon-associated hemolysin of Vibrio parahaemolyticus. Infect Immun. 1984 Jul;45(1):290–292. doi: 10.1128/iai.45.1.290-292.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kaper J. B., Levine M. M. Cloned cholera enterotoxin genes in study and prevention of cholera. Lancet. 1981 Nov 21;2(8256):1162–1163. doi: 10.1016/s0140-6736(81)90605-x. [DOI] [PubMed] [Google Scholar]
  15. Kupersztoch Y. M., Helinski D. R. A catenated DNA molecule as an intermediate in the replication of the resistance transfer factor R6K in Escherichia coli. Biochem Biophys Res Commun. 1973 Oct 15;54(4):1451–1459. doi: 10.1016/0006-291x(73)91149-2. [DOI] [PubMed] [Google Scholar]
  16. Maas R. An improved colony hybridization method with significantly increased sensitivity for detection of single genes. Plasmid. 1983 Nov;10(3):296–298. doi: 10.1016/0147-619x(83)90045-8. [DOI] [PubMed] [Google Scholar]
  17. McConnell M. M., Smith H. R., Willshaw G. A., Scotland S. M., Rowe B. Plasmids coding for heat-labile enterotoxin production isolated from Escherichia coli O78: comparison of properties. J Bacteriol. 1980 Jul;143(1):158–167. doi: 10.1128/jb.143.1.158-167.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Moseley S. L., Echeverria P., Seriwatana J., Tirapat C., Chaicumpa W., Sakuldaipeara T., Falkow S. Identification of enterotoxigenic Escherichia coli by colony hybridization using three enterotoxin gene probes. J Infect Dis. 1982 Jun;145(6):863–869. doi: 10.1093/infdis/145.6.863. [DOI] [PubMed] [Google Scholar]
  19. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  20. Snellings N. J., Johnson E. M., Baron L. S. Genetic basis of Vi antigen expression in Salmonella paratyphi C. J Bacteriol. 1977 Jul;131(1):57–62. doi: 10.1128/jb.131.1.57-62.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Snellings N. J., Johnson E. M., Kopecko D. J., Collins H. H., Baron L. S. Genetic regulation of variable Vi antigen expression in a strain of Citrobacter freundii. J Bacteriol. 1981 Feb;145(2):1010–1017. doi: 10.1128/jb.145.2.1010-1017.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  23. Totten P. A., Holmes K. K., Handsfield H. H., Knapp J. S., Perine P. L., Falkow S. DNA hybridization technique for the detection of Neisseria gonorrhoeae in men with urethritis. J Infect Dis. 1983 Sep;148(3):462–471. doi: 10.1093/infdis/148.3.462. [DOI] [PubMed] [Google Scholar]
  24. Wahl G. M., Stern M., Stark G. R. Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl-paper and rapid hybridization by using dextran sulfate. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3683–3687. doi: 10.1073/pnas.76.8.3683. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES