Based On The Imp Peptide Nucleic Acid Probe Detection Of Riemerella Anatipestifer Serotypes 1 And GyrA Peptide Nucleic Acid Synergistic Effect Of Norfloxacin Of Research

Riemerella anatipestifer(RA)is a gram-negative bacterium which is mainly responsible for the contact infectious diseases of domestic fowl like duck,goose and Turkey.In this study,the serotype 1 of RA was detected by Imp peptide nucleic acid(PNA)probe,and the enhanced antibacterial effect of gyrA peptide nucleic acid was studied.The main contents and results are as follows: 1.The study and application of Imp peptide nucleic acid probe for detection serotype 1 of RA.The 134-145 site sequence of the imp gene was selected and the PNA-imp was synthesized by using software,5-FAM in RA imp added fluorescent labeled primers,and serotype 1 RA CH-1 strain,serotype 2 RA CH-2 strain,ATCC 11845 strain and unknown serotype RA clinical isolates as the templates for PCR amplification of PNA-imp and PCR products were hybridization,the observation film is placed under a fluorescence microscope,PNA-imp and target site specific binding will show green fluorescence spots.The results showed that: in PNA-imp 6.2 ?M and 60 °C conditions can be clearly detected RA serotype 1 strains;RA serotype 2 and 6,Escherichia coli,Salmonella typhimurium,Pasteurella multocida detection was negative,with good specificity.2.The study and application of PNA-gyrA for enhanced antibacterial effectPNA-gyrA was synthesized by using software to screen the sequence of gyrA gene at site 1591-1604.Comparison of PNA-gyrA and norfloxacin quinolones on RA CH-1 strain,RA CH-2 strain,ATCC 11845 alone and together the antibacterial effect of differences,the results showed that: PNA-gyrA alone,RA CH-1 strain,RA CH-2 strain,RA strain ATCC11845(MIC)were more than 256 ?g/mL,no antibacterial effect;the interaction between PNA-gyrA and norfloxacin,RA CH-1,RA CH-2 and ATCC 11845 strain MIC of norfloxacin was respectively reduced from 8 ?g/mL,32 ?g/mL and 16 ?g/mL to 4 ?g/mL,16 ?g/mL and 8 ?g/mL,This indicated that PNA-gyrA could significantly enhance the antibacterial effect of norfloxacin.In summary,the design of peptide nucleic acid probe hybridized with imp as target gene has good specificity in detecting serum type 1 RA.The design of peptide nucleic acid with gyrA as target gene enhanced the antibacterial effect of norfloxacin.