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Volume 2, Number 3—July 1996
Synopsis

Application of Molecular Techniques
to the Diagnosis of Microsporidial Infection

Daniel P. FedorkoComments to Author  and Yasmine M. Hijazi
Author affiliations: National Institutes of Health, Bethesda, Maryland, USA

Main Article

Table 2

SSU-rRNA PCR primer pairs for the diagnosis of microsporidial infection

Primer pair (no. nucleotides) Primer designation Organisms amplified Amplicon size in base pairs Source of Target References
5'-CACCAGGTTGATTCTGCCTGAC-3' (22) V1 Enterocytozoon bieneusi 348 Biopsied tissue Duodenal aspirates 33, 41 31
5'-ACTCAGGTGTTATACTCACGTC-3' (22) EB450 Encephalitozoon hellem Cultured organisms 33
5'-GAAACTTGTCCACTCCTTACG-3' (21) EBIEF1 E.bieneusi 607 Bile 43
5'-CCATGCACCACTCCTGCCATT-3' (21) EBIER1 Duodenal aspirates
5'-TGAGAAGTAAGATGTTTAGCA-3' (21) E.hellem 547 Cultured organisms 39
5'-GTAAAAACACTCTCACACTCA-3' (21)
5'-ATGAGAAGTGATGTGTGTGCG-3' (21) Encephalitozoon cuniculi 549 Cultured organisms 39, 40
5'-TGCCATGCACTCACAGGCATC-3' (21)
5'-CACCAGGTTGATTCTGCCTGAC-3' (22) V1 Encephalitozoon intestinalis 370 Biopsied tissue 31
5'-CTCGCTCCTTTACACTCGAA-3' (20) SI500
5'-CACCAGGTTGATTCTGCCTGAC-3' (22) PMP1* E.bieneusi 250 Stool 42
5'-CCTCTCCGGAACCAAACCTG-3' (21) PMP2 E.cuniculli 268 Cultured organisms
E.intestinalis 270 Stool
E.hellem 279 Cultured organisms

*The nucleotide sequence of primer PMP1 is identical to that of primer V1.

Main Article

References
  1. Canning  EU, Lom  J, Dykova  I. The microsporidia of vertebrates. New York: Academic Press, 1986.
  2. Matsubayashi  H, Koike  T, Mikata  T, Hagiwara  S. A case of Encephalitozoon-like body infection in man. Arch Pathol. 1959;67:1817.
  3. Modigliani  R, Bories  C, Le Charpentier  Y, Salmeron  M, Messing  B, Galian  A, Diarrhoea and malabsorption in acquired immune deficiency syndrome: a study of four cases with special emphasis on opportunistic protozoan infections. Gut. 1985;26:17987. DOIPubMedGoogle Scholar
  4. Desportes  I, Le Charpentier  Y, Galian  A, Bernard  F, CochandPriollet  B, Lavergne  A, . Occurrence of a new microsporidian: Enterocytozoon bieneusi n. g., n. sp., in the enterocytes of a human patient with AIDS. J Protozool. 1985;32:2504.PubMedGoogle Scholar
  5. Weber  R, Bryan  RT, Schwartz  DA, Owen  RL. Human microsporidial infections. Clin Microbiol Rev. 1994;7:42661.PubMedGoogle Scholar
  6. Sandfort  J, Hannemann  A, Gelderblom  H, Stark  K, Owen  RL, Ruf  B. Enterocytozoon bieneusi infection in an immunocompetent patient who had acute diarrhea and who was not infected with the human immuno-deficiency virus. Clin Infect Dis. 1994;19:5146.PubMedGoogle Scholar
  7. Orenstein  JM. Microsporidiosis in the acquired immunodeficiency syndrome. J Parasitol. 1991;77:84364. DOIPubMedGoogle Scholar
  8. He  Q, Leitch  GJ, Visvesvara  GS, Wallace  S. Effects of nifedipine, metronidazole, and nitric oxide donors on spore germination and cell culture infection of the microsporidia Encephalitozoon hellem and Encephalitozoon intestinalis. Antimicrob Agents Chemother. 1996;40:17985.PubMedGoogle Scholar
  9. Kotler  DP, Orenstein  JM. Prevalence of intestinal microsporidiosis in HIV-infected individuals referred for gastroenterological evaluation. Am J Gastroenterol. 1994;89:19982002.
  10. Shadduck  JA, Greeley  E. Microsporidia and human infections. Clin Microbiol Rev. 1989;2:15865.PubMedGoogle Scholar
  11. Didier  ES, Vossbrinck  CR, Baker  MD, Rogers  LB, Bertucci  DC, Shadduck  JA. Identification and characterization of three Encephalitozoon cuniculi strains. Parasitology. 1995;111:41121. DOIPubMedGoogle Scholar
  12. Deplazes  P, Mathis  A, Baumgartner  R, Tanner  I, Weber  R. Immunologic and molecular characteristics of Encephalitozoon-like microsporidia isolated from humans and rabbits indicate that Encephalitozoon cuniculi is a zoonotic parasite. Clin Infect Dis. 1996;22:5579.PubMedGoogle Scholar
  13. Kotler  DP, Giang  TT, Garro  ML, Orenstein  JM. Light microscopic diagnosis of microsporidiosis in patients with AIDS. Am J Gastroenterol. 1994;89:5404.PubMedGoogle Scholar
  14. Field  AS, Marriott  DJ, Hing  MC. The Warthin-Starry stain in the diagnosis of small intestinal microsporidiosis in HIV-infected patients. Folia Parasitol (Praha). 1993;40:2616.PubMedGoogle Scholar
  15. Weber  R, Bryan  RT, Owen  RL, Wilcox  CM, Gorelkin  L, Visvesvara  GS. the Enteric Opportunistic Infections Working Group. Improved light-microscopic detection of microsporidia spores in stool and duodenal aspirates. N Engl J Med. 1992;326:1616.PubMedGoogle Scholar
  16. Clarridge  JE III, Karkhanis  S, Rabeneck  L, Marino  B, Foote  LW. Quantitative light microscopic detection of Enterocytozoon bieneusi in stool specimens: a longitudinal study of human immunodeficiency virus-infected microsporidiosis patients. J Clin Microbiol. 1996;34:5203.PubMedGoogle Scholar
  17. Didier  ES, Orenstein  JM, Aldras  A, Bertucci  D, Rogers  LB, Janney  FA. Comparison of three staining methods for detecting microsporidia in fluids. J Clin Microbiol. 1995;33:313845.PubMedGoogle Scholar
  18. Beckers  PJA, Derks  GJMM, van Gool  T, Rietveld  FJR, Sauerwein  RW. Encephalitozoon intestinalis-specific monoclonal antibodies for laboratory diagnosis of microsporidiosis. J Clin Microbiol. 1996;34:2825.PubMedGoogle Scholar
  19. Visvesvara  GS, Leitch  GJ, Pieniazek  NJ, da Silva  AJ, Wallace  S, Slemenda  SB, Short-term in vitro culture and molecular analysis of the microsporidian, Enterocytozoon bieneusi. J Eukaryot Microbiol. 1995;42:50610. DOIPubMedGoogle Scholar
  20. van Gool  T, Canning  EU, Gilis  H, van den Bergh Weerman  MA, Eeftinck Schattenkerk  JKM, Dankert  J. Septata intestinalis frequently isolated from stool of AIDS patients with a new cultivation method. Parasitology. 1994;109:2819. DOIPubMedGoogle Scholar
  21. Hollister  WS, Canning  EU, Willcox  A. Evidence for widespread occurence of antibodies to Encephalitozoon cuniculi (Microspora) in man provided by ELISA and other serological tests. Parasitology. 1991;102:3343. DOIPubMedGoogle Scholar
  22. Didier  ES, Kotler  DP, Dieterich  DT, Orenstein  JM, Aldras  AM, Davis  R, Serological studies in human microsporidiosis. AIDS. 1993;7(suppl 3):S811.
  23. Didier  ES, Shaddock  JA, Didier  PJ, Millichamp  N, Vossbrinck  CR. Studies on ocular microsporidia. J Protozool. 1991;38:6358.PubMedGoogle Scholar
  24. Hollister  WS, Canning  EU. An enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to Encephalitozoon cuniculi and its use in determination of infections in man. Parasitology. 1987;94:20919. DOIPubMedGoogle Scholar
  25. Bergquist  R, Morfeldt-Mansson  L, Pehrson  PO, Petrini  B, Wasserman  J. Antibody against Encephalitozoon cuniculi in Swedish homosexual men. Scand J Infect Dis. 1984;16:38991. DOIPubMedGoogle Scholar
  26. Ombrouck  C, Romestand  B, Da Costa  JC, Desportes-Livage  I, Datry  A, Coste  F, Use of crossreactive antigens of the microsporidian Glugea atherinae for the possible detection of Enterocytozoon bieneusi by western blot. Am J Trop Med Hyg. 1995;52:8993.PubMedGoogle Scholar
  27. Vossbrinck  CR, Maddox  JV, Friedman  S, Debrunner-Vossbrinck  BA, Woese  CR. Ribosomal RNA sequence suggests microsporidia are extremely ancient eukaryotes. Nature. 1987;326:41114.
  28. Schuitema  ARJ, Harstkeerl  RA, van Gool  T, Laxminarayan  R, Terpstra  WJ. Application of the polymerase chain reaction for the diagnosis of microsporidiosis. AIDS. 1993;7(suppl 3):S5761.
  29. Hartskeerl  RA, Schuitema  ARJ, deWachter  R. Secondary structure of the small subunit ribosomal RNA sequence of the microsporidium Encephalitozoon cuniculi. Nucleic Acids Res. 1993;21:1489. DOIPubMedGoogle Scholar
  30. Vossbrinck  CR, Baker  MD, Didier  ES, Debrunner-Vossbrinck  BA, Shadduck  JA. Ribosomal DNA sequences of Encephalitozoon hellem and Encephalitozoon cuniculi: species identification and phylogenetic construction. J Euk Microbiol  1993;40:354-62.
  31. Weiss  LM, Zhu  X, Cali  A, Tanowitz  HB, Wittner  M. Utility of microsporidian rRNA in diagnosis and phylogeny: a review. Folia Parasitol (Praha). 1994;41:8190.PubMedGoogle Scholar
  32. Hartskeerl  RA, van Gool  T, Schuitema  ARJ, Didier  ES, Terpstra  WJ. Genetic and immunological characterization of the microsporidian Septata intestinalis Cali, Kotler and Orenstein, 1993: reclassification to Encephalitozoon intestinalis. Parasitology. 1995;110:27785.
  33. Zhu  X, Wittner  M, Tanowitz  HB, Kotler  D, Cali  A, Weiss  LM. Small subunit rRNA sequence of Enterocytozoon bieneusi and its potential diagnostic role with use of the polymerase chain reaction. J Infect Dis. 1993;168:15705.PubMedGoogle Scholar
  34. Silveira  H, Canning  EU. Vittaforma corneae N. Comb. for the human microsporidium Nosema corneum Shadduck, Meccoli, Davis & Font, 1990 based on its ultrastructure in the liver of experimentally infected athymic mice. J Eukaryot Microbiol. 1995;42:15865. DOIPubMedGoogle Scholar
  35. Baker  MD, Vossbrinck  CR, Didier  ES, Maddox  JV, Shadduck  JA. Small subunit ribosomal DNA phylogeny of various microsporidia with emphasis on AIDS related forms. J Euk Microbiol  1995;42:564-70.
  36. Cali  A, Kotler  DP, Orenstein  JM. Septata intestinalis, n. g., n. sp., an intestinal microsporidian associated with chronic diarrhea and dissemination in AIDS patients. J Eukaryot Microbiol. 1993;40:1012. DOIPubMedGoogle Scholar
  37. Hartskeerl  RA, Schuitema  ARJ, van Gool  T, Terpstra  WJ. Genetic evidence for the occurrence of extraintestinal Enterocytozoon bieneusi infections. Nucleic Acids Res. 1993;21:4150. DOIPubMedGoogle Scholar
  38. Persing  DH. Target selection and optimization of amplification reactions. In: Persing DH, Smith TF, Tenover FC, White TJ, editors. Diagnostic molecular microbiology: principles and applications. Washington, DC: American Society for Microbiology, 1993:88-104.
  39. Visvesvara  GS, Leitch  GJ, da Silva  AJ, Croppo  GP, Moura  H, Wallace  S, . Polyclonal and monoclonal antibody and PCR-amplified small subunit rRNA identification of a microsporidian, Encephalitozoon hellem, isolated from an AIDS patient with disseminated infection. J Clin Microbiol. 1994;32:27608.PubMedGoogle Scholar
  40. de Groote  MA, Visvesvara  G, Wilson  ML, Pieniazek  NJ, Slemenda  SB, da Silva  AJ, . Polymerase chain reaction and culture confirmation of disseminated Encephalitozoon cuniculi in a patient with AIDS: successful therapy with albendazole. J Infect Dis. 1995;171:13758.PubMedGoogle Scholar
  41. Franzen  C, Müller  A, Hegener  P, Salzberger  B, Hartmann  P, Fätkenheuer  G, . Detection of microsporidia (Enterocytozoon bieneusi) in intestinal biopsy specimens from human immunodeficiency virus-infected patients by PCR. J Clin Microbiol. 1995;33:22946.PubMedGoogle Scholar
  42. Fedorko  DP, Nelson  NA, Cartwright  CP. Identification of microsporidia in stool specimens by using PCR and restriction endonucleases. J Clin Microbiol. 1995;33:173941.PubMedGoogle Scholar
  43. da Silva  AJ, Schwartz  DA, Visvesvara  GS, de Moura  H, Slemenda  SB, Pieniazek  NJ. Sensitive PCR diagnosis of infections by Enterocytozoon bieneusi (microsporidia) using primers based on the region coding for small-subunit rRNA. J Clin Microbiol. 1996;34:9867.PubMedGoogle Scholar
  44. Didier  ES, Rogers  LB, Brush  AD, Wong  S, Traina-Dorge  V, Bertucci  D. Diagnosis of disseminated microsporidian Encephalitozoon hellem infection by PCR-Southern analysis and successful treatment with albendazole and fumagillin. J Clin Microbiol. 1996;34:94752.PubMedGoogle Scholar

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