American Journal of Ophthalmology
Volume 146, Issue 5 , Pages 714-723.e1 , November 2008

Prospective Comparison of Microbial Culture and Polymerase Chain Reaction in the Diagnosis of Corneal Ulcer

  • Elma Kim

      Affiliations

    • Department of Ophthalmology and Visual Sciences, Washington University School of Medicine, St Louis, Missouri
    • ,
  • Jaya D. Chidambaram

      Affiliations

    • Francis I. Proctor Foundation, University of California, San Francisco, California
    • ,
  • Muthiah Srinivasan

      Affiliations

    • Aravind Eye Care System, Madurai, India
    • ,
  • Prajna Lalitha

      Affiliations

    • Aravind Eye Care System, Madurai, India
    • ,
  • Daniel Wee

      Affiliations

    • Department of Ophthalmology and Visual Sciences, Washington University School of Medicine, St Louis, Missouri
    • ,
  • Thomas M. Lietman

      Affiliations

    • Francis I. Proctor Foundation, University of California, San Francisco, California
    • ,
  • John P. Whitcher

      Affiliations

    • Francis I. Proctor Foundation, University of California, San Francisco, California
    • ,
  • Russell N. Van Gelder

      Affiliations

    • Department of Ophthalmology and Visual Sciences, Washington University School of Medicine, St Louis, Missouri
    • Department of Molecular Biology and Pharmacology, Washington University School of Medicine, St Louis, Missouri
    • Department of Ophthalmology, University of Washington School of Medicine, Seattle, Washington
    • Corresponding Author InformationInquiries to Russell N. Van Gelder, Department of Ophthalmology, University of Washington School of Medicine, Campus Box 356485, 1959 NE Pacific Street, Seattle, WA 98195

,Accepted 10 June 2008.

References 

  1. Whitcher JP, Srinivasan M, Upadhyay MP. Prevention of corneal ulceration in the developing world. Int Ophthalmol Clin. 2002;42:71–77
  2. Srinivasan M, Gonzales CA, George C, et al. Epidemiology and aetiological diagnosis of corneal ulceration in Madurai, South India. Br J Ophthalmol. 1997;81:965–971
  3. Upadhyay MP, Karmacharya PC, Koirala S, et al. Epidemiologic characteristics, predisposing factors, and etiologic diagnosis of corneal ulceration in Nepal. Am J Ophthalmol. 1991;111:92–99
  4. Gonzales CA, Srinivasan M, Whitcher JP, Smolin G. Incidence of corneal ulceration in Madurai district, South India. Ophthalmic Epidemiol. 1996;3:159–166
  5. Erie JC, Nevitt MP, Hodge DO, Ballard DJ. Incidence of ulcerative keratitis in a defined population from 1950 through 1988. Arch Ophthalmol. 1993;111:1665–1671
  6. Upadhyay MP, Karmacharya PC, Koirala S, et al. The Bhaktapur eye study: ocular trauma and antibiotic prophylaxis for the prevention of corneal ulceration in Nepal. Br J Ophthalmol. 2001;85:388–392
  7. Whitcher JP, Srinivasan M, Upadhyay MP. Corneal blindness: a global perspective. Bull World Health Organ. 2001;79:214–221
  8. Whitcher JP, Srinivasan M. Corneal ulceration in the developing world—a silent epidemic. Br J Ophthalmol. 1997;81:622–623
  9. Van Gelder RN. Applications of the polymerase chain reaction to diagnosis of ophthalmic disease. Surv Ophthalmol. 2001;46:248–258
  10. Farhatullah S, Kaza S, Athmanathan S, Garg P, Reddy SB, Sharma S. Diagnosis of herpes simplex virus-1 keratitis using Giemsa stain, immunofluorescence assay, and polymerase chain reaction assay on corneal scrapings. Br J Ophthalmol. 2004;88:142–144
  11. Rudolph T, Welinder-Olsson C, Lind-Brandberg L, Stenevi U. 16S rDNA PCR analysis of infectious keratitis: a case series. Acta Ophthalmol Scand. 2004;82:463–467
  12. Kumar M, Mishra NK, Shukla PK. Sensitive and rapid polymerase chain reaction based diagnosis of mycotic keratitis through single-stranded conformation polymorphism. Am J Ophthalmol. 2005;140:851–857
  13. Schabereiter-Gurtner C, Maca S, Rolleke S, et al. 16S rDNA-based identification of bacteria from conjunctival swabs by PCR and DGGE fingerprinting. Invest Ophthalmol Vis Sci. 2001;42:1164–1171
  14. Knox CM, Cevallos V, Margolis TP, Dean D. Identification of bacterial pathogens in patients with endophthalmitis by 16S ribosomal DNA typing. Am J Ophthalmol. 1999;128:511–512
  15. Schabereiter-Gurtner C, Maca S, Kaminsky S, Rolleke S, Lubitz W, Barisani-Asenbauer T. Investigation of an anaerobic microbial community associated with a corneal ulcer by denaturing gradient gel electrophoresis and 16S rDNA sequence analysis. Diagn Microbiol Infect Dis. 2002;43:193–199
  16. Gaudio PA, Gopinathan U, Sangwan V, Hughes TE. Polymerase chain reaction based detection of fungi in infected corneas. Br J Ophthalmol. 2002;86:755–760
  17. Lane DJ. 16S/23S rRNA sequencing. In:  Stackebrandt E,  Goodfellow M editor. Nucleic Acid Techniques in Bacterial Systematics. Chichester, United Kingdom: John Wiley & Sons; 1991;p. 115–175
  18. Einsele H, Hebart H, Roller G, et al. Detection and identification of fungal pathogens in blood by using molecular probes. J Clin Microbiol. 1997;35:1353–1360
  19. Rowther FB, Rodrigues C, Mehta AP, et al. An improved method of elimination of DNA from PCR reagents. Mol Diagn. 2005;9:53–57
  20. Mazzariol A, Zuliani J, Fontana R, Cornaglia G. Isolation from blood culture of a Leclercia adecarboxylata strain producing an SHV-12 extended-spectrum beta-lactamase. J Clin Microbiol. 2003;41:1738–1739
  21. Fattal O, Deville JG. Leclercia adecarboxylata peritonitis in a child receiving chronic peritoneal dialysis. Pediatr Nephrol. 2000;15:186–187
  22. Temesgen Z, Toal DR, Cockerill FR. Leclercia adecarboxylata infections: case report and review. Clin Infect Dis. 1997;25:79–81
  23. Moreira BM, Leobons MB, Pellegrino FL, et al. Ralstonia pickettii and Burkholderia cepacia complex bloodstream infections related to infusion of contaminated water for injection. J Hosp Infect. 2005;60:51–55
  24. Kismet E, Atay AA, Demirkaya E, et al. Two cases of Ralstonia pickettii bacteremias in a pediatric oncology unit requiring removal of the Port-A-Caths. J Pediatr Hematol Oncol. 2005;27:37–38
  25. Carrell DT, Emery BR, Hamilton B. Seminal infection with Ralstonia picketti and cytolysosomal spermophagy in a previously fertile man. Fertil Steril. 2003;79:1665–1667
  26. Coenye T, Vandamme P, LiPuma JJ. Infection by Ralstonia species in cystic fibrosis patients: identification of R. pickettii and R. mannitolilytica by polymerase chain reaction. Emerg Infect Dis. 2002;8:692–696
  27. Urbina BY, Gohh R, Fischer SA. Oerskovia xanthineolytica endocarditis in a renal transplant patient: case report and review of the literature. Transpl Infect Dis. 2003;5:195–198
  28. Niamut SM, van der Vorm ER, van Luyn-Wiegers CG, Gokemeijer JD. Oerskovia xanthineolytica bacteremia in an immunocompromised patient without a foreign body. Eur J Clin Microbiol Infect Dis. 2003;22:274–275
  29. Harrington RD, Lewis CG, Aslanzadeh J, Stelmach P, Woolfrey AE. Oerskovia xanthineolytica infection of a prosthetic joint: case report and review. J Clin Microbiol. 1996;34:1821–1824
  30. Shah M, Gentile RC, McCormick SA, Rogers SH. Oerskovia xanthineolytica keratitis. CLAO J. 1996;22:96
  31. Kirsch LS, Brownstein S, Deschenes J, Sorgini C, Jackson WB. Saccharomyces keratitis and endophthalmitis. Can J Ophthalmol. 1999;34:229–232
  32. Pharmakakis NM, Andrikopoulos GK, Papadopoulos GE, Petropoulos IK, Kolonitsiou FI, Koliopoulos JX. Does identification of the causal organism of corneal ulcers influence the outcome?. Eur J Ophthalmol. 2003;13:11–17
  33. Morlet N, Minassian D, Butcher J. Risk factors for treatment outcome of suspected microbial keratitis (Ofloxacin Study Group). Br J Ophthalmol. 1999;83:1027–1031
  34. Rodman RC, Spisak S, Sugar A, Meyer RF, Soong HK, Musch DC. The utility of culturing corneal ulcers in a tertiary referral center versus a general ophthalmology clinic. Ophthalmology. 1997;104:1897–1901
  35. Dahlgren MA, Lingappan A, Wilhelmus KR. The clinical diagnosis of microbial keratitis. Am J Ophthalmol. 2007;143:940–944
  36. McLeod SD, Kolahdouz-Isfahani A, Rostamian K, Flowers CW, Lee PP, McDonnell PJ. The role of smears, cultures, and antibiotic sensitivity testing in the management of suspected infectious keratitis. Ophthalmology. 1996;103:23–28
  37. Knox CM, Chandler D, Short GA, Margolis TP. Polymerase chain reaction-based assays of vitreous samples for the diagnosis of viral retinitis (Use in diagnostic dilemmas). Ophthalmology. 1998;105:37–44discussion 44–45
  38. Elnifro EM, Cooper RJ, Klapper PE, Yeo AC, Tullo AB. Multiplex polymerase chain reaction for diagnosis of viral and chlamydial keratoconjunctivitis. Invest Ophthalmol Vis Sci. 2000;41:1818–1822
  39. Crouse CA, Pflugfelder SC, Pereira I, Cleary T, Rabinowitz S, Atherton SS. Detection of herpes viral genomes in normal and diseased corneal epithelium. Curr Eye Res. 1990;9:569–581
  40. Okhravi N, Adamson P, Matheson MM, Towler HM, Lightman S. PCR-RFLP-mediated detection and speciation of bacterial species causing endophthalmitis. Invest Ophthalmol Vis Sci. 2000;41:1438–1447
  41. Knox CM, Cevellos V, Dean D. 16S ribosomal DNA typing for identification of pathogens in patients with bacterial keratitis. J Clin Microbiol. 1998;36:3492–3496
  42. Kumar M, Shukla PK. Single-stranded conformation polymorphism of large subunit of ribosomal RNA is best suited to diagnosing fungal infections and differentiating fungi at species level. Diagn Microbiol Infect Dis. 2006;56:45–51
  43. Bharathi MJ, Ramakrishnan R, Meenakshi R, Mittal S, Shivakumar C, Srinivasan M. Microbiological diagnosis of infective keratitis: comparative evaluation of direct microscopy and culture results. Br J Ophthalmol. 2006;90:1271–1276
  44. Bharathi MJ, Ramakrishnan R, Meenakshi R, Padmavathy S, Shivakumar C, Srinivasan M. Microbial keratitis in South India: influence of risk factors, climate, and geographical variation. Ophthalmic Epidemiol. 2007;14:61–69
  45. Pichare A, Patwardhan N, Damle AS, Deshmukh AB. Bacteriological and mycological study of corneal ulcers in and around Aurangabad. Indian J Pathol Microbiol. 2004;47:284–286
  46. Leck AK, Thomas PA, Hagan M, et al. Aetiology of suppurative corneal ulcers in Ghana and South India, and epidemiology of fungal keratitis. Br J Ophthalmol. 2002;86:1211–1215
  47. Couble A, Rodriguez-Nava V, de Montclos MP, Boiron P, Laurent F. Direct detection of Nocardia spp. in clinical samples by a rapid molecular method. J Clin Microbiol. 2005;43:1921–1924
  48. Ando N, Takatori K. Fungal flora of the conjunctival sac. Am J Ophthalmol. 1982;94:67–74
  49. Moeller CT, Branco BC, Yu MC, Farah ME, Santos MA, Hofling-Lima AL. Evaluation of normal ocular bacterial flora with two different culture media. Can J Ophthalmol. 2005;40:448–453
  50. Venugopal PL, Venugopal TL, Gomathi A, Ramakrishna ES, Ilavarasi S. Mycotic keratitis in Madras. Indian J Pathol Microbiol. 1989;32:190–197
  51. Valenton MJ, Rinaldi MG, Butler EE. A corneal abscess due to the fungus Botryodiplodia theobromae. Can J Ophthalmol. 1975;10:416–418
  52. Virgile R, Perry HD, Pardanani B, et al. Human infectious corneal ulcer caused by Pythium insidiosum. Cornea. 1993;12:81–83
  53. Krajaejun T, Pracharktam R, Wongwaisayawan S, Rochanawutinon M, Kunakorn M, Kunavisarut S. Ocular pythiosis: is it under-diagnosed?. Am J Ophthalmol. 2004;137:370–372
  54. Kunavisarut S, Nimvorapan T, Methasiri S. Pythium corneal ulcer in Ramathibodi Hospital. J Med Assoc Thai. 2003;86:338–342
  55. Murdoch D, Parr D. Pythium insidiosum keratitis. Aust N Z J Ophthalmol. 1997;25:177–179
  56. Badenoch PR, Coster DJ, Wetherall BL, et al. Pythium insidiosum keratitis confirmed by DNA sequence analysis. Br J Ophthalmol. 2001;85:496
  57. Carandina G, Bacchelli M, Virgili A, Strumia R. Simonsiella filaments isolated from erosive lesions of the human oral cavity. J Clin Microbiol. 1984;19:931–933
  58. Okhravi N, Adamson P, Carroll N, et al. PCR-based evidence of bacterial involvement in eyes with suspected intraocular infection. Invest Ophthalmol Vis Sci. 2000;41:3474–3479
  59. Welinder-Olsson C, Dotevall L, Hogevik H, et al. Comparison of broad-range bacterial PCR and culture of cerebrospinal fluid for diagnosis of community-acquired bacterial meningitis. Clin Microbiol Infect. 2007;13:879–886
  60. Dempsey KE, Riggio MP, Lennon A, et al. Identification of bacteria on the surface of clinically infected and non-infected prosthetic hip joints removed during revision arthroplasties by 16S rRNA gene sequencing and by microbiological culture. Arthritis Res Ther. 2007;9:R46
  61. Mardh PA, Novikova N, Witkin SS, Korneeva I, Rodrigues AR. Detection of candida by polymerase chain reaction vs microscopy and culture in women diagnosed as recurrent vulvovaginal cases. Int J STD AIDS. 2003;14:753–756
  62. Ryan MP, Pembroke JT, Adley CC. Ralstonia pickettii: a persistent gram-negative nosocomial infectious organism. J Hosp Infect. 2006;62:278–284
  63. De Baere T, Wauters G, Huylenbroeck A, et al. Isolations of Leclercia adecarboxylata from a patient with a chronically inflamed gallbladder and from a patient with sepsis without focus. J Clin Microbiol. 2001;39:1674–1675
  64. Borst A, Box AT, Fluit AC. False-positive results and contamination in nucleic acid amplification assays: suggestions for a prevent and destroy strategy. Eur J Clin Microbiol Infect Dis. 2004;23:289–299
  65. Padua RA, Parrado A, Larghero J, Chomienne C. UV and clean air result in contamination-free PCR. Leukemia. 1999;13:1898–1899
  66. Fung DYC, Crozier-Dodson BA. Recovery and identification of airborne microorganisms in swine facilities using selective agar and thin agar layer resuscitation media. http://www.pork.org/PorkScience/Research/Documents/02-023-FUNG.3-31-03.pdfAccessed: June 6, 2008
  67. Birenzvige A, Eversole J, Seaver M, Francesconi S, Valdes E, Kulaga H. Aeorosol characteristics in a subway environment. Aerosol Sci Technol. 2003;37:210–220
  68. Loeffler J, Hebart H, Bialek R, et al. Contaminations occurring in fungal PCR assays. J Clin Microbiol. 1999;37:1200–1202
  69. Rimek D, Garg AP, Haas WH, Kappe R. Identification of contaminating fungal DNA sequences in Zymolyase. J Clin Microbiol. 1999;37:830–831
  70. Muyzer G, de Waal EC, Uitterlinden AG. Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes coding for 16S rRNA. Appl Environ Microbiol. 1993;59:695–700
  71. Sharma S, Kunimoto DY, Gopinathan U, Athmanathan S, Garg P, Rao GN. Evaluation of corneal scraping smear examination methods in the diagnosis of bacterial and fungal keratitis: a survey of eight years of laboratory experience. Cornea. 2002;21:643–647

 Supplemental Material available at AJO.com.

PII: S0002-9394(08)00473-X

doi: 10.1016/j.ajo.2008.06.009

American Journal of Ophthalmology
Volume 146, Issue 5 , Pages 714-723.e1 , November 2008

Article Tools

* Image Usage & Resolution