ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2017, Vol. 48 ›› Issue (3): 538-543.doi: 10.11843/j.issn.0366-6964.2017.03.018

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Real-time Fluorescence Loop-mediated Isothermal Amplification for Detection of hCD39 in Transgenic Pig

LI Jin1，LIU Wei1，LI Jun-chao2，JIN Hong-yan1，LI Gang1*   

  1. (1. Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China; 2. Department of Cardiovascular, China PLA General Hospital, Beijing 100853, China)
  • Received:2016-08-30 Online:2017-03-23 Published:2017-03-23

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Abstract:

Using ESE-Quant tube scanner testing platform, we established a real-time fluorescence loop-mediated isothermal amplification (RTF-LAMP) method for rapid detection of hCD39. The method employed a set of six specially designed primers for amplification of nucleic acid under isothermal conditions at 63 ℃ for 45 min. The amplification process of LAMP was monitored by the addition of SYBR Green Ⅰ dye prior to amplification. The fluorescence intensity of SYBR Green Ⅰ dye was used as the standard of the results. Results of the study showed that the sensitivity of the method was 3 fg•μL－1, and there was no cross-reactivity with the genes of related and unrelated detected, such as hCD46, LEA29Y. More importantly, this method could be completed testing of buccal swabs specimens of transgenic pigs. The purpose of our study was to develop a simple, economical, rapid method with good sensitivity and specificity. The method could be no damage to the living body samples for testing, suitable for quick and easy on-site operation.

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