Use of laser capture microdissection together with in situ hybridization and real-time PCR to study distribution of latent herpes simplex virus genomes in mouse trigeminal ganglia.
The herpes simplex virus (HSV) frequently establishes a latent state in neurons, which can then be reactivated from the infected neurons. Quantifying the single-cell viral load is essential for understanding latency and reactivation of this virus. In this chapter the methods of laser capture microdissection and quantitative real-time polymerase chain reaction with in situ hybridization have been combined to determine the HSV copy number per neuron in latently infected trigeminal ganglia. The distribution of latent herpes simplex genomes at the individual cell level has been deteremined and the relationship of the number of latent genomes to the expression of latency-associated transcripts established.
Chen
Xiao-Ping
XP
Department of Microbiology and Immunology, Tong-Ji University School of Medicine, Shanghai, China.
Mata
Marina
M
Fink
David J
DJ
D000818:Animals; D005260:Female; D016679:Genome, Viral; D006561:Herpes Simplex / Q000235:genetics / Q000473:pathology / Q000821:virology; D017403:In Situ Hybridization / Q000379:methods*; D007834:Lasers; D051379:Mice; D008807:Mice, Inbred BALB C; D042282:Microdissection / Q000379:methods*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D018139:Simplexvirus / Q000235:genetics* / Q000502:physiology; D012668:Trigeminal Ganglion / Q000473:pathology / Q000821:virology*; D017735:Virus Latency