1887

Microbiology Society logo

Volume 150, Issue 5

Molecular typing of found in formalin-fixed paraffin-embedded lung tissue sections from sudden infant death victims Free

Abstract

Previous studies have provided histological evidence of an association between primary infection and sudden infant death syndrome (SIDS). The aim of this work was to determine the species of clustered organisms found in formalin-fixed paraffin-embedded (FFPE) lung tissue sections from Chilean sudden infant death (SID) victims. This approach needed first to optimize a DNA extraction method from such histological sections. For that purpose, the QIAamp DNA Isolation from Paraffin-Embedded Tissue method (Qiagen) was first tested on FFPE lung tissue sections of immunosuppressed Wistar rats inoculated with rat-derived . Successful DNA extraction was assessed by the amplification of a 346 bp fragment of the mitochondrial large subunit rRNA gene of the species using a previously described PCR assay. PCR products were analysed by direct sequencing and sequences corresponding to were found in all the samples. This method was then applied to FFPE lung tissue sections from Chilean SID victims. was successfully identified in the three tested samples. In conclusion, an efficient protocol for isolating PCR-ready DNA from FFPE lung tissue sections was developed. It established that the species found in the lungs of Chilean SID victims was .

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
Keyword(s): FFPE, formalin-fixed paraffin-embedded , mt LSU, mitochondrial large subunit , SID, sudden infant death and SIDS, sudden infant death syndrome
SGM
Loading

Article metrics loading...

/content/journal/micro/10.1099/mic.0.26895-0
2004-05-01
2024-04-18
Loading full text...

Full text loading...

/deliver/fulltext/micro/150/5/mic1501167.html?itemId=/content/journal/micro/10.1099/mic.0.26895-0&mimeType=html&fmt=ahah

References

  1. Aliouat E. M., Dei-Cas E., Ouaissi A., Palluault F., Soulez B., Camus D. 1993; In vitro attachment of Pneumocystis carinii from mouse and rat origin. Biol Cell 77:209–217 [CrossRef]
     [Google Scholar]
  2. Aviles P., Aliouat E. M., Martinez A., Dei-Cas E., Herreros E., Dujardin L., Gargallo-Viola D. 2000; In vitro pharmacodynamic parameters of sordarin derivatives in comparison with those of marketed compounds against Pneumocystis carinii isolated from rats. Antimicrob Agents Chemother 44:1284–1290 [CrossRef]
     [Google Scholar]
  3. Chabé M., Dei-Cas E., Creusy C., Fleurisse L., Respaldiza N., Camus D., Durand-Joly I. 2004; Immunocompetent hosts as a reservoir of Pneumocystis organisms: histological and RT-PCR data demonstrate active replication. Eur J Clin Microbiol Infect Dis 23:89–97 [CrossRef]
     [Google Scholar]
  4. Dei-Cas E., Brun-Pascaud M., Bille-Hansen V., Allaert A., Aliouat E. M. 1998; Animal models of pneumocystosis. FEMS Immunol Med Microbiol 22:163–168 [CrossRef]
     [Google Scholar]
  5. Dumoulin A., Mazars E., Seguy N., Gargallo-Viola D., Vargas S., Cailliez J. C., Aliouat E. M., Wakefield A. E., Dei-Cas E. 2000; Transmission of Pneumocystis carinii disease from immunocompetent contacts of infected hosts to susceptible hosts. Eur J Clin Microbiol Infect Dis 19:671–678 [CrossRef]
     [Google Scholar]
  6. Durand-Joly I., Aliouat el M., Recourt C., Guyot K., Francois N., Wauquier M., Camus D., Dei-Cas E. 2002; Pneumocystis cariniif. sp. hominis is not infectious for SCID mice. J Clin Microbiol 40:1862–1865 [CrossRef]
     [Google Scholar]
  7. Durand-Joly I., Soula F., Chabé M., Dalle J. H., Lafitte J. J., Sénéchal M., Pinon A., Camus D., Dei-Cas E. 2003; Long-term colonization with Pneumocystis jirovecii in hospital staffs: a challenge to prevent nosocomial pneumocystosis. J Eukaryot Microbiol 50 : Suppl 614–615
     [Google Scholar]
  8. ECA 1996; In vitro systems in Pneumocystis research: the members of the European concerted action on Pneumocystis. Parasitol Today 12:245–249 [CrossRef]
     [Google Scholar]
  9. Frank T. S., Svoboda-Newman S. M., Hsi E. D. 1996; Comparison of methods for extracting DNA from formalin-fixed paraffin sections for nonisotopic PCR. Diagn Mol Pathol 5:220–224 [CrossRef]
     [Google Scholar]
  10. Frenkel J. K. 1999; Pneumocystis pneumonia, an immunodeficiency-dependent disease (IDD): a critical historical overview. J Eukaryot Microbiol 46:89S–92S
     [Google Scholar]
  11. Goelz S. E., Hamilton S. R., Vogelstein B. 1985; Purification of DNA from formaldehyde fixed and paraffin embedded human tissue. Biochem Biophys Res Commun 130:118–126 [CrossRef]
     [Google Scholar]
  12. Greer C. E., Peterson S. L., Kiviat N. B., Manos M. M. 1991; PCR amplification from paraffin-embedded tissues. Effects of fixative and fixation time. Am J Clin Pathol 95:117–124
     [Google Scholar]
  13. Lane B. R., Ast J. C., Hossler P. A., Mindell D. P., Bartlett M. S., Smith J. W., Meshnick S. R. 1997; Dihydropteroate synthase polymorphisms in Pneumocystis carinii. J Infect Dis 175:482–485 [CrossRef]
     [Google Scholar]
  14. Latouche S., Ortona E., Mazars E., Margutti P., Tamburrini E., Siracusano A., Guyot K., Nigou M., Roux P. 1997; Biodiversity of Pneumocystis carinii hominis: typing with different DNA regions. J Clin Microbiol 35:383–387
     [Google Scholar]
  15. Lee C. H., Lu J. J., Bartlett M. S., Durkin M. M., Liu T. H., Wang J., Jiang B., Smith J. W. 1993; Nucleotide sequence variation in Pneumocystis carinii strains that infect humans. J Clin Microbiol 31:754–757
     [Google Scholar]
  16. Meuwissen J. H., Tauber I., Leeuwenberg A. D., Beckers P. J., Sieben M. 1977; Parasitologic and serologic observations of infection with Pneumocystis in humans. J Infect Dis 136:43–49 [CrossRef]
     [Google Scholar]
  17. Morgan D. J., Vargas S. L., Reyes-Mugica M., Walterspiel J. N., Carver W., Gigliotti F. 2001; Identification of Pneumocystis carinii in the lungs of infants dying of sudden infant death syndrome. Pediatr Infect Dis J 20:306–309 [CrossRef]
     [Google Scholar]
  18. Nevez G., Totet A., Pautard J. C., Raccurt C. 2001; Pneumocystis carinii detection using nested-PCR in nasopharyngeal aspirates of immunocompetent infants with bronchiolitis. J Eukaryot Microbiol Suppl122S–123S
     [Google Scholar]
  19. Peglow S. L., Smulian A. G., Linke M. J.7 other authors 1990; Serologic responses to Pneumocystis carinii antigens in health and disease. J Infect Dis 161:296–306 [CrossRef]
     [Google Scholar]
  20. Pifer L. L., Hughes W. T., Stagno S., Woods D. 1978; Pneumocystis carinii infection: evidence for high prevalence in normal and immunosuppressed children. Pediatrics 61:35–41
     [Google Scholar]
  21. Sheldon W. H. 1959; Subclinical pneumocystis pneumonitis. AMA J Dis Child 97:287–297
     [Google Scholar]
  22. Sinclair K., Wakefield A. E., Banerji S., Hopkin J. M. 1991; Pneumocystis carinii organisms derived from rat and human hosts are genetically distinct. Mol Biochem Parasitol 45:183–184 [CrossRef]
     [Google Scholar]
  23. Totet A., Latouche S., Lacube P., Bolognini J., Raccurt C., Nevez G., Roux P. 2001; Typing of Pneumocystis carinii f.sp. hominis isolates from nasopharyngeal aspirates of immunocompetent infants with bronchiolitis by dihydropteroate synthase gene analysis. J Eukaryot Microbiol Suppl 121S:
     [Google Scholar]
  24. Totet A., Pautard J. C., Raccurt C., Roux P., Nevez G. 2003a; Genotypes at the internal transcribed spacers of the nuclear rRNA operon of Pneumocystis jiroveci in nonimmunosuppressed infants without severe pneumonia. J Clin Microbiol 41:1173–1180 [CrossRef]
     [Google Scholar]
  25. Totet A., Respaldiza N., Pautard J. C., Raccurt C., Nevez G. 2003b; Pneumocystis jiroveci genotypes and primary infection. Clin Infect Dis 36:1340–1342 [CrossRef]
     [Google Scholar]
  26. Tsolaki A. G., Miller R. F., Underwood A. P., Banerji S., Wakefield A. E. 1996; Genetic diversity at the internal transcribed spacer regions of the rRNA operon among isolates of Pneumocystis carinii from AIDS patients with recurrent pneumonia. J Infect Dis 174:141–156 [CrossRef]
     [Google Scholar]
  27. Tsolaki A. G., Beckers P., Wakefield A. E. 1998; Pre-AIDS era isolates of Pneumocystis cariniif. sp. hominis: high genotype similarity with contemporary isolates. J Clin Microbiol 36:90–93
     [Google Scholar]
  28. Vargas S. L., Ponce C. A., Hughes W. T.12 other authors 1999; Association of primary Pneumocystis carinii infection and sudden infant death syndrome. Clin Infect Dis 29:1489–1493 [CrossRef]
     [Google Scholar]
  29. Vargas S. L., Hughes W. T., Santolaya M. E., Ulloa A. V., Ponce C. A., Cabrera C. E., Cumsille F., Gigliotti F. 2001; Search for primary infection by Pneumocystis carinii in a cohort of normal, healthy infants. Clin Infect Dis 32:855–861 [CrossRef]
     [Google Scholar]
  30. Wakefield A. E. 1996; DNA sequences identical to Pneumocystis carinii f. sp. carinii and Pneumocystis cariniif. sp. hominis in samples of air spora. J Clin Microbiol 34:1754–1759
     [Google Scholar]
  31. Wakefield A. E. 1998; Genetic heterogeneity in human-derived Pneumocystis carinii. FEMS Immunol Med Microbiol 22:59–65 [CrossRef]
     [Google Scholar]
  32. Wakefield A. E., Stewart T. J., Moxon E. R., Marsh K., Hopkin J. M. 1990a; Infection with Pneumocystis carinii is prevalent in healthy Gambian children. Trans R Soc Trop Med Hyg 84:800–802 [CrossRef]
     [Google Scholar]
  33. Wakefield A. E., Pixley F. J., Banerji S., Sinclair K., Miller R. F., Moxon E. R., Hopkin J. M. 1990b; Detection of Pneumocystis carinii with DNA amplification. Lancet 336:451–453 [CrossRef]
     [Google Scholar]
  34. Wakefield A. E., Fritscher C. C., Malin A. S., Gwanzura L., Hughes W. T., Miller R. F. 1994; Genetic diversity in human-derived Pneumocystis carinii isolates from four geographical locations shown by analysis of mitochondrial rRNA gene sequences. J Clin Microbiol 32:2959–2961
     [Google Scholar]
  35. Wu L., Patten N., Yamashiro C. T., Chui B. 2002; Extraction and amplification of DNA from formalin-fixed, paraffin-embedded tissues. Appl Immunohistochem Mol Morphol 10:269–274
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/mic.0.26895-0
Loading
Molecular typing of Pneumocystis jirovecii found in formalin-fixed paraffin-embedded lung tissue sections from sudden infant death victims
Microbiology 150, 1167 (2004); https://doi.org/10.1099/mic.0.26895-0
/content/journal/micro/10.1099/mic.0.26895-0
/content/journal/micro/10.1099/mic.0.26895-0
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error