In situ hybridization detection of varicella zoster virus in paraffin-embedded skin biopsy samples.
BACKGROUND
When virologic and molecular diagnostic techniques are unavailable, the diagnosis of varicella zoster virus (VZV) infection depends on clinical criteria and histologic evaluation of skin biopsy specimens or Tzank preparations. These methods can misdiagnose chickenpox and zoster, particularly when the clinical manifestations are atypical.


OBJECTIVE
To improve diagnosis in these settings, we developed an in situ hybridization technique for the detection of VZV utilizing a fluorescein-labeled oligonucleotide probe visualized with anti-fluorescein alkaline phosphatase-conjugated antibody.


METHODS
We retrospectively examined 26 paraffin-embedded skin biopsy specimens with histologic features consistent with VZV or herpes simplex virus (HSV) infection and 11 control cases by in situ hybridization. In situ hybridization for VZV and HSV-1 was compared with polymerase chain reaction (PCR) for VZV and HSV-1 and clinical and histologic examination.


RESULTS
Thirteen of the 26 study cases and two of the 11 control cases were positive for VZV by in situ hybridization. When compared with PCR, in situ hybridization was 92% sensitive and 88% specific. When compared with clinical diagnosis, in situ hybridization was 86% sensitive and 87% specific. All cases of chickenpox had VZV-positive inflammatory cells in the dermis but this finding was less frequent among the cases of zoster.


CONCLUSIONS
This in situ hybridization technique is a sensitive and specific method for the diagnosis of VZV in skin lesions that is applicable to most histopathology laboratory settings. In addition, in situ hybridization reveals individual infected cells and may provide insight into the pathogenesis of VZV skin infection.
Annunziato
P
P
Department of Pediatrics, Columbia University, College of Physicians and Surgeons, New York, NY 10032, USA.
Lungu
O
O
Gershon
A
A
Silvers
D N
DN
LaRussa
P
P
Silverstein
S J
SJ
D002644:Chickenpox / Q000473:pathology / Q000821:virology; D004279:DNA, Viral / Q000032:analysis; D006562:Herpes Zoster / Q000473:pathology / Q000821:virology; D018259:Herpesvirus 1, Human / Q000235:genetics / Q000302:isolation & purification; D014645:Herpesvirus 3, Human / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction; D012189:Retrospective Studies; D012680:Sensitivity and Specificity; D012867:Skin / Q000473:pathology / Q000821:virology*
D004279:DNA, Viral