Localization of Herpes Simplex Virus Type 1 DNA in Latently Infected BALB/c Mice Neurons Using in situ Polymerase Chain Reaction

Authors
Behzad Khansarinejad
Hoorieh Soleimanjahi
Amir Ghaemi
Taki Tiraihi
Shahram Pour Beiranvand
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Abstract
Background: Herpes simplex virus type-1 (HSV-1) establishes a lifelong latent infection in neurons following primary infection. The existence of latent HSV-1 DNA in the trigeminal ganglia of infected BALB/c mice was examined using a direct in situ PCR technique, based on Digoxigenin-11-dUTP detection system with anti-digoxigenin-peroxidase and 3,3'-diaminobenzidine (DAB) substrate. Methods: Eight-week-old male BALB/c mice were inoculated via the eye by 104 plaque forming unit of wild type Iranian isolates of HSV-1. After establishment of latency, trigeminal ganglia were removed and examined using in situ PCR to detect HSV-1 genome. Finally, the results of in situ PCR were verified by a two-round PCR method, using amplification cocktail of in situ reaction, as a template for a conventional gel base PCR. Results and Conclusion: The results suggest that a direct in situ PCR method using a peroxidase and DAB detection system is a useful means for detection of latent HSV-1 DNA in the latently infected ganglia.
Keywords
Herpes simplex virus-1
Latency
In situ PCR
two-round PCR
Trigeminal ganglia