A non-radioactive method for identifying enzyme-amplified products of the reticuloendotheliosis proviral env and LTR genes using psoralen-biotin labelled probes.

Abstract

A novel polymerase chain reaction (PCR) system based on the env gene of reticuloendotheliosis virus (REV) strain REV-A for the detection of proviral DNA is described. The designed PCR product of 807 bp was identified using an internal probe of 278 bp produced by nested PCR from REV-infected DNA CEF. The env-gene PCR was then compared with the previously described PCR for proviral REV-long terminal repeat and the PCR product served also as the probe. The probes were labelled with the psoralen-biotin system by photoactivation and the southern blot hybridization signal was detected colorimetricaly. The advantages of using a non-radioactive means of probe labelling were demonstrated clearly in that study, as well as the effective labeling of probes with psoralen-biotin and the simple colorimetric method of detection. The env-gene PCR detected all eleven REV strains used in the study. These included three REV prototype strains and eight Israeli REV isolates. Both PCR systems had similar levels of sensitivity.

• Authors: I Davidson, M Malkinson
• Journal: Journal of virological methods (J Virol Methods) Vol. 59 Issue 1-2 Pg. 113-9 (May 1996) ISSN: 0166-0934 [Print] Netherlands
• PMID: 8793837 (Publication Type: Comparative Study, Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• DNA Probes
• DNA, Viral
• Gene Products, env
• Biotin
• Ficusin

Topics

• Animals
• Biotin
• Chick Embryo
• Chickens
• DNA Probes
• DNA, Viral (analysis)
• Ducks
• Ficusin
• Gene Products, env (genetics)
• Polymerase Chain Reaction
• Proviruses (genetics, isolation & purification)
• Repetitive Sequences, Nucleic Acid
• Reticuloendotheliosis virus (genetics, isolation & purification)
• Sensitivity and Specificity
• Turkey