Current Microbiology

, Volume 52, Issue 5, pp 379–382 | Cite as

Evaluation of Herpes Simplex Detection in Corneal Scrapings by Three Molecular Methods

  • Amina Mostafa Abd El-Aal
  • Maysaa El Sayed
  • Eglal Mohammed
  • Mohammed Ahmed
  • Mona Fathy
Article

Abstract

Herpes simplex virus (HSV)-1 keratitis (HSK) is a sight-threatening ocular infection with worldwide occurrence. A prompt laboratory diagnosis is often very useful. The purpose of this study was to evaluate molecular methods as rapid diagnostic tools compared with cell culture of HSK. Corneal scrapings from patients with clinically suspected HSK were tested by direct immunofluorescence assay (IFA) for HSV-1 antigen and by polymerase chain reaction (PCR) for HSV-1 DNA, and an attempt for viral isolation was performed on Vero cell line culture. Positive samples by cell culture were 20.8%, whereas PCR was positive in 29.2%, and IFA was positive in 33.3%. IFA had better sensitivity (80%) and negative predictive value (81.8%) than PCR (70% and 76.9%, respectively); however, PCR had better specificity (71.4%) and positive predictive value (63.6%). This indicates that a combination of cell culture, IFA and PCR constitutes the best set of tools for diagnosis of clinically suspected cases of HSK. Documented infection can be further assessed by cell-culture technique or PCR depending laboratory availability.

Literature Cited

  1. 1.
    Butler TK, Spencer NA, Chan CC, Singh Gilhortra J, McClella K (2005) Infective keratitis in older patients: A 4 year review, 1998–2002. Br J Ophthalmol 89:591–596CrossRefPubMedGoogle Scholar
  2. 2.
    Athmanathan S, Bandlapally SR, Rao GN (2001) Collection of corneal impression cytology directly on a sterile glass slide for the detection of viral antigen an inexpensive and simple technique for the diagnosis of HSV epithelial keratitis—A pilot study. BMC Ophthalmol. 1:3–10PubMedGoogle Scholar
  3. 3.
    Madhavan HN, Priya K, Anand AR, Therese KL (1999). Detection of herpes simplex (HSV) genome using polymerase chain reaction (PCR) in clinical samples comparison of PCR with standard laboratory methods for detection of HSV. J Clin Virol 14:145–151CrossRefPubMedGoogle Scholar
  4. 4.
    Kowalski RP, Gordon YJ (1989). Evaluation of immunologic tests for the detection of ocular herpes simplex virus. Ophthalmology 96:1583–1586PubMedGoogle Scholar
  5. 5.
    Thiel MA, Bossart W, Bernauer W (1997). Improved impression cytology techniques for the immunopathological diagnosis of superficial viral infections. Br J Ophthalmol 81:984–988PubMedGoogle Scholar
  6. 6.
    Pramod NP, Gopalakrishnan V, Mohan R, Chandriga S, Anadakannan K, Menon T, et al. (1998). Enhanced detection of herpes simplex virus from ocular specimens of herpetic keratitis patients. Indian J Pathol Microbiol 41:49–53PubMedGoogle Scholar
  7. 7.
    Pramod NP, Thyagarajan SP, Mohan KV, Anandakannan K (2000). Polymerase chain reaction in the diagnosis of herpetic keratitis: Experience in a developing country. Can J Ophthalmol 35:134–140PubMedGoogle Scholar
  8. 8.
    Khodadoost MA, Sabahi F, Behroz MJ, Roustai MH, Saderi H, Amini-Bavil-Olyaee S, et al. (2004). Study of a polymerase chain reaction based method for detection of herpes simplex virus 1 DNA among Iranian patients with ocular herpetic keratitis infection. Jpn J Ophthalmol 48:328–332CrossRefPubMedGoogle Scholar
  9. 9.
    Lucotte G, Bathelier C, Lespiaux V, Bali C, Champenios T (1995). Detection and genotyping of herpes simplex virus types 1 and 2 by polymerase chain reaction. Mol Cell Probes 9:287–290CrossRefPubMedGoogle Scholar
  10. 10.
    Whiley DM, Mackay IM, Syrmis MW, Witt MJ, Sloots TP (2004). Detection and differentiation of herpes simplex virus type 1 and 2 by a duplex light cycler PCR that incorporates an internal control PCR reaction. J Clin Virol 30:32–38CrossRefPubMedGoogle Scholar
  11. 11.
    Schmidt NJ (1979). Cell culture techniques for diagnostic virology. In: Lennette EH, Schmidt NJ (eds) Diagnostic procedures for viral, richettsial and chlamydial infections. Washington, DC: American Public Health Association, pp 65–139Google Scholar
  12. 12.
    Landry ML, Hsuing GD (2001) Primary isolation of viruses. In: Specter S, Hodinka RL, Young SA (eds) Clinical virology manual, 3rd ed, ASM Press, Washington, pp 27–42Google Scholar
  13. 13.
    Johnson G, Nelson S, Petric M, Tellier R (2000). Comprehensive PCR-based assay for detection and species identification of human herpes viruses. J Clin Microb 38:3274–3279Google Scholar
  14. 14.
    Biriken D, Yildiz S, Ozkul A, Ozsan M (2004). Investigation of herpes simplex type 1 in the samples of patients with clinically prediagnosed as herpetic keratitis or keratoconjunctivitis. Mikrobiyol Bul 38:51–59PubMedGoogle Scholar
  15. 15.
    Elnifro EM, Cooper RJ, Klapper PE, et al. (1999). Diagnosis of viral and chlamydial keratoconjunctivitis : Which laboratory test? Br J Ophthalmol 83:622–627PubMedGoogle Scholar
  16. 16.
    Farhatullah S, Kaja S, Athmanathan S, Reddy SB, Sharma S (2004). Diagnosis of herpes simplex virus-1 keratitis using Giemsa stain, immunofluorescence assay and polymerase chain reaction assay on corneal scrapings. Br J Ophthalmol 88:142–144CrossRefPubMedGoogle Scholar
  17. 17.
    Kodama T, Hayasaka S, Setogawa T (1992). Immunofluorescence staining and corneal sensitivity in patients suspected of having herpes simplex keratitis . Am J Ophthalmol 113:187–89PubMedGoogle Scholar
  18. 18.
    Kaye SB, Lynas C, Patterson A (2000). Human herpes viruses in the cornea. Br J Ophthalmol 84:563–571CrossRefPubMedGoogle Scholar
  19. 19.
    Chan EL, Brandt K, Horsman GB (2001). Comparison of chemicon simulfluor direct fluorescent antibody staining with cell culture and shell vial direct immunoperoxidase staining for detection of herpes simplex virus and direct immunofluorescence staining for detection of varicella zoster virus. Clin Diagn Lab Immunol 8:909–912CrossRefPubMedGoogle Scholar
  20. 20.
    Subhan S, Jose RJ, Duggirala A, et al. (2004). Diagnosis of herpes simplex virus-1 keratitis: Comparison of Giemsa, immunofluorescence assay and polymerase chain reaction. Curr Eye Res 29:209–113CrossRefPubMedGoogle Scholar
  21. 21.
    Kowalski RP, Gordon YJ, Romanowski EG, et al. (1993) A comparison of enzyme immunoassay and polymerase chain reaction with the clinical examination for diagnosing ocular herpetic disease. Ophthalmology 100:530–534PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, Inc. 2006

Authors and Affiliations

  • Amina Mostafa Abd El-Aal
    • 1
  • Maysaa El Sayed
    • 1
  • Eglal Mohammed
    • 2
  • Mohammed Ahmed
    • 3
  • Mona Fathy
    • 1
  1. 1.Department of Clinical Pathology, Faculty of MedicineMansoura UniversityEgypt
  2. 2.Department of Ophthalmology, Faculty of MedicineMansoura UniversityEgypt
  3. 3.National Research CenterEgypt

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