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Studies on the mycoplasmal pneumonia in slaughter pigs. 2. Isolation of mycoplasmas from lung tissues and identification of isolates by nested-PCR technique  

Lim, Young-Taek (Department of Animal Science, College of life Sciences, Dankook University)
Seok, Ho-Bong (Department of Animal Science, College of life Sciences, Dankook University)
Publication Information
Korean Journal of Veterinary Research / v.42, no.2, 2002 , pp. 225-229 More about this Journal
Abstract
We report that mycoplasma organisms from lung tissues of slaughter pigs were identified to genes fragments with references use of nested-PCR technique(nPCR). Seven strains of mycoplasma species were isolated from 70 lung tissues. The organisms were detected by in vitro amplification of 16S rRNA and 23S rRNA genes. Nucleotide sequences of the spacer between 16S and 23S in the ribosomal RNA operons of mycoplasma were identified by the analysis of products from the nested PCR. Four common PCR primers, MhF1, MhF2 MhR1 and MhR2, were designed by analysis between these sequences by first amplified with F1, R1 and second with F2, R2, respectively. Specific amplification of the spacer region for reference strains of M. hyopneumoniae, M. hyorhinis, M. flocculare were confirmed by first round of PCR in which the traduced fragments of 690bp, 460bp, 630bp. But amplications of second round was changed to 240bp, 210bp, 230bp, respectively. Three different strains (M. hyopneumoniae:4, M. hyorhinis:2, M. flocculare:1) were detected by the nested-PCR technique. The results suggest that the detection of swine mycoplasma by n-PCR can be analyzed the nucleotide sequences between rRNA operons and homology study.
Keywords
mycoplamsal pneumonia; slaughter pigs; Mycoplasma species; lung tissues; nested-PCR;
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