Elia Gabriella, Decaro Nicola, Martella Vito, Campolo Marco, Desario Costantina, Lorusso Eleonora, Cirone Francesco, Buonavoglia Canio
Department of Animal Health and Well-being, Faculty of Veterinary Medicine of Bari, 70010 Valenzano, Bari, Italy.
J Virol Methods. 2006 Apr;133(1):70-5. doi: 10.1016/j.jviromet.2005.10.024. Epub 2005 Nov 22.
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.
开发了一种用于检测和定量1型马疱疹病毒(EHV-1)的实时PCR检测方法。将该检测方法的灵敏度与已建立的巢式PCR(n-PCR)进行了比较。实时PCR能检测到1个拷贝的靶DNA,其灵敏度比基于凝胶的n-PCR高1个对数级。该检测方法能够特异性地检测马组织样本中的EHV-1 DNA,且不会对其他马疱疹病毒进行交叉扩增。应用实时PCR来测定马流产胎儿组织样本中的EHV-1载量。EHV-1特异性荧光PCR检测方法具有高灵敏度和可重复性,结合宽动态范围和高通量,使其适用于诊断和研究应用。