›› 2012, Vol. 39 ›› Issue (12): 24-26.

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Detection of Mycoplasma mycoides Cluster by Loop-mediated Isothermal Amplification

XIE Xiu-lan1, KANG Xiao-dong1, CHU Yue-feng2, MA Xiao-ming1, YUE Cai-juan1   

  1. 1. Breeding Engineering Technology Center,Ningxia Academy of Agriculture and Forestry Sciences,Yinchuan 750002, China;
    2. Key Laboratory of Grazing Animal Diseases of Ministry of Agriculture, Lanzhou Veterinary Research Institute,Lanzhou 730046, China
  • Received:2012-06-14 Online:2012-12-20 Published:2012-12-19

Abstract: In this study, a sensitive loop-mediated isothermal amplification (LAMP) was developed for rapidly detecting Mycoplasma mycoides cluster. A set of four primers,two outer and two inner primers, were designed from genomic sequences according to the conserved region of the 16S rRNA gene of Mycoplasma mycoides cluster. The reaction was performed in one step in a single tube at 65 ℃ for 60 min, with hydroxynaphthol blue (HNB) dye added prior to amplification. Blue-positive results were significantly different from the purple-negative. The assay could amplify from Mycoplasma mycoides cluster members, Mmc, Mmm LC and Mccp,but no cross-reaction from other bacteria. The sensitivity test showed that it could detect 10 pg/μL DNA from Mmc strain PG3. The study had developed a diagnostic procedure which was rapid and sensitive for Mycoplasma mycoides cluster.

Key words: Mycoplasma mycoides cluster; LAMP; diagnosis

CLC Number: