Development of a real-time PCR for the detection and quantitation of caprine herpesvirus 1 in goats.
Caprine herpesvirus 1 (CpHV-1) is an alphaherpesvirus interfering with goat reproductive performances. The virus is associated with neonatal mortality in kids and reproductive failure in adults. A real-time PCR assay based on TaqMan technology and targeting the gene encoding for glycoprotein C (gC) was developed for detection and quantitation of CpHV-1 in samples collected from infected goats. The detection limit of the assay was 1 x 10(2) standard DNA copies, with a sensitivity of 1-2 logs higher than the conventional gel-based PCR assay targeting the same gene. The real-time PCR was reproducible, as shown by satisfactory low intra-assay and interassay coefficients of variation. The quantitative assay was validated on clinical samples, including genital swabs and various tissue samples collected from goats either infected naturally or experimentally with CpHV-1. The high sensitivity, simplicity and reproducibility of the CpHV-1 fluorogenic PCR assay, combined with its wide dynamic range and high throughput, make this method especially suitable for studies on the pathogenesis and for trials with experimental vaccines and antiviral drugs.
Elia
Gabriella
G
Department of Animal Health and Well-being, Faculty of Veterinary Medicine, University of Bari, Sp Casamassima km. 3, 70010 Valenzano, Italy.
Tarsitano
Elvira
E
Camero
Michele
M
Bellacicco
Anna Lucia
AL
Buonavoglia
Domenico
D
Campolo
Marco
M
Decaro
Nicola
N
Thiry
Julien
J
Tempesta
Maria
M
D000818:Animals; D015511:Goat Diseases / Q000821:virology*; D006041:Goats; D006566:Herpesviridae Infections / Q000662:veterinary / Q000821:virology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D018140:Varicellovirus / Q000235:genetics* / Q000302:isolation & purification*; D014764:Viral Proteins / Q000235:genetics
D014764:Viral Proteins