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IBJ-Iranian Biomedical Journal. 2010; 14 (3): 83-88
in English | IMEMR | ID: emr-108582

ABSTRACT

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 3,3f-diaminobenzidine [DAB] substrate. Eight-week-old male BALB/c mice were inoculated via the eye by 10[4] 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. 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


Subject(s)
Male , Animals, Laboratory , Polymerase Chain Reaction , Trigeminal Ganglion/virology , Virus Latency , DNA, Viral , Mice, Inbred BALB C , Neurons/virology , Digoxigenin/analogs & derivatives , Deoxyuracil Nucleotides
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