Key result
A one-tube multiplex PCR method was developed to unambiguously discriminate between ev1+/ev1+ homozygote, ev1-/ev1- homozygote, and ev1+/ev1- heterozygote chickens.
A one-tube multiplex PCR method was developed to rapidly and safely detect the endogenous Rous-associated virus element ev1 in chickens, suitable for large-scale screening.
New multiplex PCR enables rapid ev1 detection in chickens; leaves open validation for breeding programs.
A quick and simple method has been developed to detect the presence or absence of the endogenous Rous-associated virus (RAV) element ev1 in chickens. The procedure consists of a one-tube multiplex polymerase chain reaction (PCR) involving three oligonucleotide primers that are specific for the upstream flanking region, the long terminal repeat (LTR), and the downstream flanking region of the proviral insert, respectively. The multiplex reaction allows for the unambiguous discrimination between ev1+/ev1+ homozygote, ev1-/ev1- homozygote, and ev1+/ev1- heterozygote birds. The method works best with purified genomic DNA as substrate, but can also be used with rapidly prepared, "crude" DNA samples. The combination of speed with the safety of a nonradioactive procedure, and the ability to perform large numbers of assays by a semi-automated procedure, make this method attractive for large-scale screening projects. The ev1 locus has been used as a model system to demonstrate the feasibility of the PCR diagnostic approach. However the same principle should be applicable to the analysis of other RAV-type ev loci, as well as endogenous elements belonging to other families of viruses as sequence information for the flanking regions of these inserts becomes available.
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Benkel et al. (1992) studied Endogenous Rous-associated virus (RAV) element ev1 in chickens. One-tube multiplex polymerase chain reaction (PCR) was evaluated on Detection of the presence or absence of the endogenous Rous-associated virus (RAV) element ev1. A one-tube multiplex PCR method was developed to unambiguously discriminate between ev1+/ev1+ homozygote, ev1-/ev1- homozygote, and ev1+/ev1- heterozygote chickens.
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