A nested PCR for the detection and differentiation of EHV-1 and EHV-4.
The nested PCR method was applied for the detection and direct differentiation of equine herpesvirus type 1 (EHV-1) and type 4 (EHV-4). Primer pairs were chosen from the glycoprotein B (gB) coding region of each serotype. The outer and inner EHV-1 primer pairs were type-specific, whereas the outer EHV-4 primer pair amplified EHV-1 and EHV-4 DNA and was therefore suitable for the detection of both virus types in a single sample. However, the nested EHV-4 primer pair was type-specific. The advantages of the nested PCR are twofold. Firstly, this assay was in the case of EHV-1 100 times, and in the case of EHV-4 1000 times more sensitive than the standard PCR, which indicates a detection limit of 1-0.1 fg of DNA (3-1 genome equivalents). Secondly, it allows the direct differentiation of EHV-1 and -4 without the need to resort to further analytical techniques.
Borchers
K
K
Institut für Virologie der Freien Universität Berlin, Germany.
Slater
J
J
D018138:Alphaherpesvirinae / Q000235:genetics / Q000302:isolation & purification*; D001483:Base Sequence; D017931:DNA Primers; D004279:DNA, Viral / Q000235:genetics; D005814:Genes, Viral / Q000235:genetics; D006023:Glycoproteins / Q000235:genetics; D004861:Herpesvirus 1, Equid / Q000235:genetics / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D017931:DNA Primers; D004279:DNA, Viral; D006023:Glycoproteins