The Establishment Of Detection Method For Mycoplasma Ovipneumoniae And Analysis Of It's Genome Bioinformatics

The study aimed at construction of a prompt TaqMan real-time fluorescent quantitation PCR method for detection of Mycoplasma ovipneumoniae,and performed whole genome sequencing of the MO HHHT2017 strain to find its biological information.In the study,the primer and the probe were disigned by the gene sequence of HSP70 which had been published in the GenBank,and the standard plasmid was recombinanted.The results showed that the linearity of standard curve was good:R~2=1.000,Slope=-3.334,E=99.5%.The sensitivity of the TaqMan real-time fluorescent quantitation PCR was 100times higher than the regular PCR,and the specificity was good for identifying Pasteurella?Streptococcus and Mycoplasma bovis etc.In addition,the coefficient of variation of Ct values was all lower than 2%no matter within or between groups.Using the TaqMan real-time fluorescent quantitation PCR could identify the Mycoplasma ovipneumoniae with less time and more accuracy.The results of the research would provide technical support for the diagnosis,prevention and control of Mycoplasmal pneumonia of sheep.The analasis to the genome of HHHT2017 showed that the whole genome size of HHHT2017 was 1019689 bp,the content of GC was 29.068%,and it has 807 genes which represented 86.87%of the full genome.The COG,GO annotation and the CRISPR prediction were carried out to the whole genome of HHHT2017.30 tRNAs and 3 rRNAs were predicted in the whole genome.In virulence gene prediction,there were 5 virulence genes which had a similarity of more than 80%with the genes published in the database.The key biological information of Mycoplasma ovipneumoniae was found in the study and the information would lay the foundation for the subsequent development of vaccines against the pathogen.