Infectious bronchitis virus: detection and vaccine Strain differentiation by semi-nested RT-PCR

摘要: A semi-nested reverse transcription-polymerase chain reaction (Semi- N-RT-PCR) was developed and used to detect the S glycoprotein gene of infectious bronchitis virus (IBV) strains discriminate H120 vaccine strain from other strains. Viral RNA extracted allantoic fluid chicken embryos tissues chickens experimentally infected with different IBV. Amplification identification viral performed using two sets primers complementary a region in Semi-N-RT-PCR assay. The pair first PCR consisted universal oligonucleotides flanking more variable S1-S2 gene. second primer comprised one together either another internal oligolucleotide or oligonucleotide sequence specific for detected all reference IBV field isolates tested herein. Semi-N-RT- had high sensitivity specificity, able differentiate strains; including M41 strain. All tissue samples collected were positive RT-PCR primers, while only H120-infected amplified by set containing H120-oligonucleotide. In conclusion, ability distinct preliminarily (H120) closes diagnostic gap offers opportunity use comprehensive procedures diagnosis.