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. 2010 Jul;14(3):83-8.

Localization of herpes simplex virus type 1 DNA in latently infected BALB/c mice neurons using in situ polymerase chain reaction

Behzad Khansarinejad  1 , Hoorieh Soleimanjahi  1 , Amir Ghaemi  1 , Taki Tiraihi  2 , Shahram Pour Beiranvand  2
Affiliations

Localization of herpes simplex virus type 1 DNA in latently infected BALB/c mice neurons using in situ polymerase chain reaction

Behzad Khansarinejad et al. Iran Biomed J. 2010 Jul.

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.

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