Detection of equine herpesvirus type 1 by real time PCR.
A real-time PCR assay was developed for detection and quantitation of equid herpesvirus type 1 (EHV-1). The sensitivity of the assay was compared with an established nested-PCR (n-PCR). The real-time PCR detected 1 copy of target DNA, with a sensitivity 1 log higher than gel-based n-PCR. The assay was able to detect specifically EHV-1 DNA in equine tissue samples and there was no cross-amplification of other horse herpesviruses. Real-time PCR was applied to determine EHV-1 load in tissue samples from equine aborted fetuses. The high sensitivity and reproducibility of the EHV-1-specific fluorogenic PCR assay, combined with the wide dynamic range and the high throughput, make this method suitable for diagnostic and research applications.
Elia
Gabriella
G
Department of Animal Health and Well-being, Faculty of Veterinary Medicine of Bari, 70010 Valenzano, Bari, Italy. g.elia@veterinaria.uniba.it
Decaro
Nicola
N
Martella
Vito
V
Campolo
Marco
M
Desario
Costantina
C
Lorusso
Eleonora
E
Cirone
Francesco
F
Buonavoglia
Canio
C
D037881:Aborted Fetus / Q000821:virology; D000818:Animals; D004279:DNA, Viral / Q000032:analysis* / Q000235:genetics; D005260:Female; D018628:Gene Dosage; D006566:Herpesviridae Infections / Q000662:veterinary* / Q000821:virology; D004861:Herpesvirus 1, Equid / Q000235:genetics*; D006734:Horse Diseases / Q000821:virology*; D006736:Horses; D011247:Pregnancy; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary*; D012680:Sensitivity and Specificity; D019562:Viral Load
D004279:DNA, Viral