title: Evaluation of tear samples for Herpes Simplex Virus 1 (HSV) detection in suspected cases of viral keratitis using PCR assay and conventional laboratory diagnostic tools.
abstract: BACKGROUND
  Herpes Simplex Virus (HSV) keratitis is a leading cause of corneal blindness. Definitive laboratory diagnosis is essential for timely management. Collection of corneal scrapings in patients with advanced epithelial keratitis and corneal thinning poses perforation risks; tear fluid is a feasible and convenient alternative but has not been widely evaluated for HSV detection.
  
  
  METHODS
  Tear fluid alone (229) or along with corneal scrapings (153) from patients of suspected herpetic keratitis was tested for HSV-1 antigen by indirect immunofluorescence assay, virus isolation in Hep 2 cells and PCR to amplify the 111bp region of the thymidine kinase (tk) coding gene and the 144bp region from the DNA polymerase coding gene of HSV.
  
  
  RESULTS
  HSV 1 antigen was detected in 31/229 (13.53%) tear specimen and 35/153 (22.87%) corneal scrapings in immunofluorescence assay; virus was isolated from 12/229 (5.2%) tear and 17/153 (11.11%) corneal scrapings, and PCR was positive for both the genes in 32/229 (13.97%) tear specimen and 56/153 (36.66%) corneal scrapings.
  
  
  CONCLUSIONS
  Corneal scrapings yielded a significantly better HSV positivity than tears in both the PCR assay (p<0.0005) and immunofluorescence assay. PCR was much more sensitive than immunofluorescence and virus isolation. However, tears should be tested for definitive laboratory diagnosis of HSV infection whenever corneal scraping collection is not possible.
authors.affiliation: Department of Ocular Microbiology, Dr R P Center for Ophthalmic Sciences, All India Institute of Medical Sciences, Ansari Nagar, New Delhi 110029, India. gita.satpathy@gmail.com
authors.forename: Gita
authors.initials: G
authors.lastname: Satpathy
authors.forename: Abhishek Kumar
authors.initials: AK
authors.lastname: Mishra
authors.forename: Radhika
authors.initials: R
authors.lastname: Tandon
authors.forename: Manoj Kumar
authors.initials: MK
authors.lastname: Sharma
authors.forename: Anjana
authors.initials: A
authors.lastname: Sharma
authors.forename: Niranjan
authors.initials: N
authors.lastname: Nayak
authors.forename: Jeewan Singh
authors.initials: JS
authors.lastname: Titiyal
authors.forename: Namrata
authors.initials: N
authors.lastname: Sharma
mesh_terms: D000956:Antigens, Viral / Q000032:analysis; D004279:DNA, Viral / Q000032:analysis; D005260:Female; D005455:Fluorescent Antibody Technique; D018259:Herpesvirus 1, Human / Q000276:immunology / Q000302:isolation & purification*; D006801:Humans; D016849:Keratitis, Herpetic / Q000175:diagnosis* / Q000276:immunology / Q000821:virology; D008297:Male; D016133:Polymerase Chain Reaction; D013666:Tears / Q000276:immunology / Q000821:virology*
chemical_list: D000956:Antigens, Viral; D004279:DNA, Viral