›› 2010, Vol. 37 ›› Issue (6): 164-167.

• 疾病防治 • Previous Articles     Next Articles

Development of the Method for Detection and Quantitation of Leishmania with Real-time Fluorescent Quantitative PCR

ZHANG Rui-yan1，SHANG Li-min1，ZHANG Ning2，LIU Quan1，WU Yong-kui1   

  1. (1. Military Veterinary Institute，Academy of Military Sciences of PLA，Changchun 130062，China； 2. College of Animal Science and Veterinary Medicine，Jilin University，Changchun 130062，China)
  • Received:1900-01-01 Revised:2010-03-13 Online:2010-06-20 Published:2010-06-20
  • Contact: LIU Quan

Abstract: To establish a SYBR-Green I fluorescent quantitative PCR method to detect the minicircles of the kinetoplast DNA of Leishmania，special primers were designed and synthesized to this conserved sequence. The special fragment was cloned into pMD18-T vector after PCR amplification，and then was transformed into E.coli DH5α. After the positive recombinant plasmid was identified，it was used as quantitative template to generate standard curve and melt curve. Results showed that the standard curve had a good linear relationship between cycle threshold(Ct) and template concentration，and the correlation coefficient was 0.996. We have successfully established a SYBR-Green I fluorescent quantitative PCR method to detect Leishmania，which can be used in the diagnosis and epidemiological surveillance of Leishmaniasis.

Key words: Leishmania; real-time fluorescent quantitative PCR; kDNA

CLC Number: