›› 2015, Vol. 42 ›› Issue (4): 859-864.doi: 10.16431/j.cnki.1671-7236.2015.04.013

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Development of a SYBR Green Ⅰ Real-time PCR Method for Detection of Chicken Infectious Anemia Virus

YANG Li-cong1,2, SU Xia2, ZHU Rui-hao2, ZHOU Hong-zhuan2, XU Fu-zhou2, YANG Bing2, SUN Ji-guo1   

  1. 1. College of Veterinary Medicine, Agricultural University of Hebei, Baoding 071000, China;
    2. Beijing Key Laboratory for Prevention and Control of Infectious Diseases in Livestock and Poultry, Institute of Animal Science and Veterinary Medicine, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097, China
  • Received:2014-11-21 Online:2015-04-20 Published:2015-05-05

Abstract: The aim of this study was to develop a Real-time PCR method for detection of chicken infectious anemia virus (CIAV). According to the CIAV genome conservative area, a pair of specific primers which could amplify about 180 bp fragment was designed. By building the pGM-T-CIAV recombinant plasmid, and preparing positive standard, the SYBR GreenⅠReal-time PCR standard curve was established. At the same time, sensitivity test, specificity test and repeatability test were determined. The results indicated that CIAV Ct threshold had good linear relationship with the standard concentration. The correlation rate and slope were 0.998 and -3.443, respectively. The Tm was about 86 ℃. No cross-reactions were found between REV, ALV J subtype, MDV and IBDV. The sensibility was 5.33 coppies/μL, which was 1 000 times than the traditional PCR. The coefficient of variation value was less than 3%. The CIAV SYBR Green Ⅰ Real-time PCR detection method could be realized on the early diagnosis of the disease and infection degree quantitative analysis of the test.

Key words: chicken infectious anemia virus (CIAV); Real-time PCR; SYBR Green Ⅰ; quantitative analysis

CLC Number: