Abstract
The purpose of this study was to identify the most useful gene for the detection of biodiversity of Pneumocystis carinii hominis isolates and to compare samples from French and Italian subjects. We studied 20 bronchoalveolar lavage fluid specimens from 20 human immunodeficiency virus-infected patients (10 French and 10 Italian patients) with Pneumocystis carinii pneumonia by DNA sequencing of the thymidylate synthase (TS), 5S rRNA, large-subunit mitochondrial rRNA (mt LSU rRNA), and internal transcribed spacer (ITS1 and ITS2) genes. Thirteen of the 20 sequenced samples had the prototype TS gene sequence. Fourteen of the 20 samples showed the prototype sequence of the 5S rRNA gene, and 6 had variant sequences of the 5S rRNA gene. The mt LSU rRNA gene was sequenced for 18 of the 20 samples; all sequences were different from the prototype sequence and were classified into four groups. Thirteen of the 20 ITS1 and ITS2 sequences were analyzed, and all the sequences were found to be different from the prototype sequence and were classified into 10 groups. The internal transcribed spacer regions thus appear to be the most discriminatory region of DNA for analysis of the biodiversity of P. carinii hominis isolates.
Full Text
The Full Text of this article is available as a PDF (195.9 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bauer N. L., Paulsrud J. R., Bartlett M. S., Smith J. W., Wilde C. E., 3rd Immunologic comparisons of Pneumocystis carinii strains obtained from rats, ferrets, and mice using convalescent sera from the same sources. J Protozool. 1991 Nov-Dec;38(6):166S–168S. [PubMed] [Google Scholar]
- Edlind T. D., Bartlett M. S., Weinberg G. A., Prah G. N., Smith J. W. The beta-tubulin gene from rat and human isolates of Pneumocystis carinii. Mol Microbiol. 1992 Nov;6(22):3365–3373. doi: 10.1111/j.1365-2958.1992.tb02204.x. [DOI] [PubMed] [Google Scholar]
- Edman U., Edman J. C., Lundgren B., Santi D. V. Isolation and expression of the Pneumocystis carinii thymidylate synthase gene. Proc Natl Acad Sci U S A. 1989 Sep;86(17):6503–6507. doi: 10.1073/pnas.86.17.6503. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hong S. T., Steele P. E., Cushion M. T., Walzer P. D., Stringer S. L., Stringer J. R. Pneumocystis carinii karyotypes. J Clin Microbiol. 1990 Aug;28(8):1785–1795. doi: 10.1128/jcm.28.8.1785-1795.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Keely S. P., Stringer J. R., Baughman R. P., Linke M. J., Walzer P. D., Smulian A. G. Genetic variation among Pneumocystis carinii hominis isolates in recurrent pneumocystosis. J Infect Dis. 1995 Aug;172(2):595–598. doi: 10.1093/infdis/172.2.595. [DOI] [PubMed] [Google Scholar]
- Kitada K., Oka S., Kimura S., Shimada K., Serikawa T., Yamada J., Tsunoo H., Egawa K., Nakamura Y. Detection of Pneumocystis carinii sequences by polymerase chain reaction: animal models and clinical application to noninvasive specimens. J Clin Microbiol. 1991 Sep;29(9):1985–1990. doi: 10.1128/jcm.29.9.1985-1990.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kovacs J. A., Halpern J. L., Lundgren B., Swan J. C., Parrillo J. E., Masur H. Monoclonal antibodies to Pneumocystis carinii: identification of specific antigens and characterization of antigenic differences between rat and human isolates. J Infect Dis. 1989 Jan;159(1):60–70. doi: 10.1093/infdis/159.1.60. [DOI] [PubMed] [Google Scholar]
- Latouche S., Roux P., Poirot J. L., Lavrard I., Hermelin B., Bertrand V. Preliminary results of Pneumocystis carinii strain differentiation by using molecular biology. J Clin Microbiol. 1994 Dec;32(12):3052–3053. doi: 10.1128/jcm.32.12.3052-3053.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lee C. H., Lu J. J., Bartlett M. S., Durkin M. M., Liu T. H., Wang J., Jiang B., Smith J. W. Nucleotide sequence variation in Pneumocystis carinii strains that infect humans. J Clin Microbiol. 1993 Mar;31(3):754–757. doi: 10.1128/jcm.31.3.754-757.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lu J. J., Bartlett M. S., Shaw M. M., Queener S. F., Smith J. W., Ortiz-Rivera M., Leibowitz M. J., Lee C. H. Typing of Pneumocystis carinii strains that infect humans based on nucleotide sequence variations of internal transcribed spacers of rRNA genes. J Clin Microbiol. 1994 Dec;32(12):2904–2912. doi: 10.1128/jcm.32.12.2904-2912.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mazars E., Odberg-Ferragut C., Dei-Cas E., Fourmaux M. N., Aliouat E. M., Brun-Pascaud M., Mougeot G., Camus D. Polymorphism of the thymidylate synthase gene of Pneumocystis carinii from different host species. J Eukaryot Microbiol. 1995 Jan-Feb;42(1):26–32. doi: 10.1111/j.1550-7408.1995.tb01536.x. [DOI] [PubMed] [Google Scholar]
- Mazars E., Odberg-Ferragut C., Durand I., Tibayrenc M., Dei-Cas E., Camus D. Genomic and isoenzymatic markers of Pneumocystis from different host species. J Eukaryot Microbiol. 1994 Sep-Oct;41(5):104S–104S. [PubMed] [Google Scholar]
- Ortona E., Margutti P., De Luca A., Tamburrini E., Visconti E., Siracusano A. Genetic variability in different Pneumocystis isolates from man and rats. New Microbiol. 1995 Jul;18(3):335–340. [PubMed] [Google Scholar]
- Perry K. M., Fauman E. B., Finer-Moore J. S., Montfort W. R., Maley G. F., Maley F., Stroud R. M. Plastic adaptation toward mutations in proteins: structural comparison of thymidylate synthases. Proteins. 1990;8(4):315–333. doi: 10.1002/prot.340080406. [DOI] [PubMed] [Google Scholar]
- Sinclair K., Wakefield A. E., Banerji S., Hopkin J. M. Pneumocystis carinii organisms derived from rat and human hosts are genetically distinct. Mol Biochem Parasitol. 1991 Mar;45(1):183–184. doi: 10.1016/0166-6851(91)90042-5. [DOI] [PubMed] [Google Scholar]
- Smulian A. G., Sullivan D. W., Linke M. J., Halsey N. A., Quinn T. C., MacPhail A. P., Hernandez-Avila M. A., Hong S. T., Walzer P. D. Geographic variation in the humoral response to Pneumocystis carinii. J Infect Dis. 1993 May;167(5):1243–1247. doi: 10.1093/infdis/167.5.1243. [DOI] [PubMed] [Google Scholar]
- Wakefield A. E., Fritscher C. C., Malin A. S., Gwanzura L., Hughes W. T., Miller R. F. Genetic diversity in human-derived Pneumocystis carinii isolates from four geographical locations shown by analysis of mitochondrial rRNA gene sequences. J Clin Microbiol. 1994 Dec;32(12):2959–2961. doi: 10.1128/jcm.32.12.2959-2961.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wakefield A. E., Pixley F. J., Banerji S., Sinclair K., Miller R. F., Moxon E. R., Hopkin J. M. Detection of Pneumocystis carinii with DNA amplification. Lancet. 1990 Aug 25;336(8713):451–453. doi: 10.1016/0140-6736(90)92008-6. [DOI] [PubMed] [Google Scholar]
- Yoganathan T., Lin H., Buck G. A. An electrophoretic karyotype and assignment of ribosomal genes to resolved chromosomes of Pneumocystis carinii. Mol Microbiol. 1989 Nov;3(11):1473–1480. doi: 10.1111/j.1365-2958.1989.tb00132.x. [DOI] [PubMed] [Google Scholar]