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.
Full text
PDFImages in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]
- 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]