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• W1986775239 abstract "A nested reverse transcription (RT)-polymerase chain reaction with subsequent restriction endonuclease analysis was developed for identification of the σ C-encoded gene of avian reoviruses (ARV). PCR products derived from the σ C-encoded gene of all tested ARVs resulted in a specific DNA band of 1023 bp, indicating that there were no apparent insertions or deletions in this region. Amplification with the nested primer pairs S1M-S1N and S1P-S1N generated 330 and 239 bp, respectively. PCR products amplified from the σ C-encoded of all tested ARVs isolates were further confirmed by Southern blot hybridization and restriction endonuclease analysis. PCR amplified cDNA fragment (1023 bp) cleaved with Pst I generated two fragments of 565 and 458 bp. The amplified σ C-encoded gene of ARV was subcloned into PQE 32 vector for further study of its antigenicity and immunogenicity. The sensitivity of RT-PCR was examined on nucleic acids from the ARV infected cell cultures. The detection limit was 100 to 10−1 TCID50 of ARV in a ethidium bromide stained gel and could be increased further to 10−1 to 10−2 TCID50 of ARV by Southern blot hybridization using a digoxigenin-labeled cDNA probe. The sensitivity increased approximately 103 to 104 folds when the cDNA was reamplified with two sets of nested primers." @default.
• W1986775239 created "2016-06-24" @default.
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• W1986775239 date "1999-08-01" @default.
• W1986775239 modified "2023-09-24" @default.
• W1986775239 title "Identification of the σ C-encoded gene of avian reovirus by nested PCR and restriction endonuclease analysis" @default.
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• W1986775239 doi "https://doi.org/10.1016/s0166-0934(99)00063-4" @default.
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