title: Multiplex polymerase chain reaction for the detection of herpes simplex virus, varicella-zoster virus and cytomegalovirus in ocular specimens.
abstract: OBJECTIVE
  A majority of ocular viral diseases are caused by herpes group of viruses. Such infections, especially atypical herpetic keratitis, iridocyclitis and intra-ocular inflammations, can often present with overlapping clinical manifestations misleading the diagnosis. Molecular techniques are most useful in such instances for an accurate and rapid diagnosis since conventional methods are time consuming and less sensitive. A multiplex PCR was developed and used for the detection of herpes simplex virus (HSV), varicella zoster virus (VZV), and cytomegalovirus (CMV) in ocular samples.
  
  
  METHODS
  One hundred and forty six ocular samples (corneal scrapings - 52, aqueous fluid - 36, vitreous fluid - 31, tissues - 26, skin vesicle scraping - 1) were included in the study. The sensitivity of the assay was determined using serial dilutions of standard strains of HSV, VZV, and CMV vis-à-vis plaque forming assay.
  
  
  RESULTS
  The sensitivity of the assay was 4, 4 and 12 PFU/ml or 20, 20 and 60 genome copy numbers of HSV, VZV and CMV respectively. Using DNA from various sources (fungal, bacterial, human leukocytes, tissues) along with standard positive controls, the assay was found to be highly specific. HSV DNA was detected in majority of the clinical samples (33.6%), most frequent being corneal samples. Comparatively, VZV and CMV infections were detected in small number of samples (VZV-3, CMV-2).
  
  
  CONCLUSIONS
  We found the assay very useful in our set-up whenever a differential diagnosis of herpetic infections was suggested by the ophthalmologist. The multiplex PCR we have described here can be of greater value in clinics with larger number of patients suspected of having HSV, VZV or CMV infections.
authors: Jhaveri Microbiology Center, Hyderabad Eye Research Foundation, L.V. Prasad Eye Institute, L.V Prasad Marg, Banjara Hills, Hyderabad, India.
authors: Gurunadh Reddy
authors: GR
authors: Chichili
authors: Sreedharan
authors: S
authors: Athmanathan
authors: Syed
authors: S
authors: Farhatullah
authors: Nibaran
authors: N
authors: Gangopadhyay
authors: Subhadra
authors: S
authors: Jalali
authors: Gunisha
authors: G
authors: Pasricha
authors: Savitri
authors: S
authors: Sharma
mesh_terms: D003587:Cytomegalovirus / Q000235:genetics* / Q000302:isolation & purification; D004279:DNA, Viral / Q000032:analysis; D015828:Eye Infections, Viral / Q000821:virology*; D018259:Herpesvirus 1, Human / Q000235:genetics* / Q000302:isolation & purification; D018258:Herpesvirus 2, Human / Q000235:genetics* / Q000302:isolation & purification; D014645:Herpesvirus 3, Human / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D004279:DNA, Viral