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. 1991 Dec;57(12):3666–3670. doi: 10.1128/aem.57.12.3666-3670.1991

Development of a 16S rRNA-based oligomer probe specific for Listeria monocytogenes.

R F Wang 1, W W Cao 1, M G Johnson 1
PMCID: PMC184033  PMID: 1723868

Abstract

Crude rRNA was isolated from Listeria monocytogenes, L. innocua, and L. ivanovii and sequenced by a reverse transcriptase method. Only two sequence regions were found to differ for L. monocytogenes versus L. innocua or L. ivanovii. Two oligonucleotide probes (RL-1 and RL-2) complementary to these two regions of rRNA of L. monocytogenes were synthesized. The RL-1 probe had one base while the RL-2 probe had two bases which differed for L. monocytogenes versus L. innocua and L. ivanovii. Use of a dried gel hybridization in place of Northern (RNA) hybridization or dot blot hybridization indicated that the RL-2 probe hybridized with all 36 strains of L. monocytogenes tested but not with 6 other Listeria species and 11 other bacteria tested. The RL-2 probe is specific for L. monocytogenes, while the RL-1 probe showed some cross-reactions with other Listeria species. An alkaline phosphatase-labeled RL-2 probe could be used in a dot blot hybridization test and gave good results, but a 32P-labeled RL-2 probe was more sensitive than the nonradioactive probe and the 32P-labeled probe was useable for up to 2 months, even though the 32P was highly degenerated.

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Selected References

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  1. Ahmad I., Finkelstein J. A., Steggles A. W. The analysis of RNA by in situ agarose gel hybridization is more sensitive than the equivalent northern blot analysis. Biotechniques. 1990 Feb;8(2):162–165. [PubMed] [Google Scholar]
  2. Chenevert J., Mengaud J., Gormley E., Cossart P. A DNA probe specific for L. monocytogenes in the genus Listeria. Int J Food Microbiol. 1989 Jul;8(4):317–319. doi: 10.1016/0168-1605(89)90002-0. [DOI] [PubMed] [Google Scholar]
  3. Datta A. R., Wentz B. A., Russell J. Cloning of the listeriolysin O gene and development of specific gene probes for Listeria monocytogenes. Appl Environ Microbiol. 1990 Dec;56(12):3874–3877. doi: 10.1128/aem.56.12.3874-3877.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. DeLong E. F., Wickham G. S., Pace N. R. Phylogenetic stains: ribosomal RNA-based probes for the identification of single cells. Science. 1989 Mar 10;243(4896):1360–1363. doi: 10.1126/science.2466341. [DOI] [PubMed] [Google Scholar]
  5. Giovannoni S. J., DeLong E. F., Olsen G. J., Pace N. R. Phylogenetic group-specific oligodeoxynucleotide probes for identification of single microbial cells. J Bacteriol. 1988 Feb;170(2):720–726. doi: 10.1128/jb.170.2.720-726.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Kaufmann S. H. Listeriosis: new findings--current concern. Microb Pathog. 1988 Oct;5(4):225–231. doi: 10.1016/0882-4010(88)90094-0. [DOI] [PubMed] [Google Scholar]
  7. King W., Raposa S., Warshaw J., Johnson A., Halbert D., Klinger J. D. A new colorimetric nucleic acid hybridization assay for Listeria in foods. Int J Food Microbiol. 1989 Jun;8(3):225–232. doi: 10.1016/0168-1605(89)90017-2. [DOI] [PubMed] [Google Scholar]
  8. Lamont R. J., Postlethwaite R., MacGowan A. P. Listeria monocytogenes and its role in human infection. J Infect. 1988 Jul;17(1):7–28. doi: 10.1016/s0163-4453(88)92236-0. [DOI] [PubMed] [Google Scholar]
  9. Lane D. J., Field K. G., Olsen G. J., Pace N. R. Reverse transcriptase sequencing of ribosomal RNA for phylogenetic analysis. Methods Enzymol. 1988;167:138–144. doi: 10.1016/0076-6879(88)67015-7. [DOI] [PubMed] [Google Scholar]
  10. Rosenberg H. F., Ackerman S. J., Tenen D. G. An alternative method for labeling oligonucleotide probes for screening cDNA libraries. Biotechniques. 1990 Apr;8(4):384–384. [PubMed] [Google Scholar]
  11. Wang R. F., Cao W. W., Johnson M. G. Dried gel hybridization in place of southern hybridization for detection of Listeria monocytogenes DNA fragments. Lett Appl Microbiol. 1991 Jun;12(6):224–227. doi: 10.1111/j.1472-765x.1991.tb00545.x. [DOI] [PubMed] [Google Scholar]
  12. Wang R. F., Cao W. W., Slavik M. F. Comparison and modification of rRNA sequencing methods. Biotechniques. 1991 Oct;11(4):438–440. [PubMed] [Google Scholar]
  13. Wilson K. H., Blitchington R., Hindenach B., Greene R. C. Species-specific oligonucleotide probes for rRNA of Clostridium difficile and related species. J Clin Microbiol. 1988 Dec;26(12):2484–2488. doi: 10.1128/jcm.26.12.2484-2488.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]

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