›› 2013, Vol. 40 ›› Issue (4): 55-58.

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Establishment of a Nested PCR Assay for Detection of Chicken Infectious Anemia Virus

WANG Jing-ru, YANG Xiao-jing   

  1. Liaoning Chaoyang Engineering Technical School,Chaoyang 122000,China
  • Received:2012-09-21 Online:2013-04-20 Published:2013-04-19

Abstract: According to the sequence of chicken infectious anemia virus (CIAV) strain published in GenBank,two pairs of primers were designed and synthesized.The outer primers amplified a fragment of 485 bp in length, and the inner primers amplification fragment size was 297 bp in length.A nested PCR assay for rapid detection of ALV was established.A specific 297 bp fragment was amplified from DNA templates of CAV strain,but no bands were amplified with templates extracted respectively from avian influenza virus (AIV) subtype H9,Newcastle disease virus (NDV), infectious bursal disease virus(IBDV),egg drop syndrome virus (EDSV), reticuloendotheliosis (REV), avian reovirus (ARV) and Marek’s disease virus(MDV). Sensitivity of the 1st and 2nd amplifications by the nested PCR assay were 100 pg and 1 fg,respectively.The sensitivite of the 2nd amplifications increased by 105 times.The results showed that the nested PCR was specific,sensitive,rapid,and accurate,and could be used as a routine assay for the detection of CAV.This method had good reproducibility, specificity and sensitivity, and might detect low content CAV accurately and rapidly.This method could be used as a method for the diagnosis and detection of clinical cases,and molecular epidemiological investigation of CAV.

Key words: chicken infectious anemia virus; nested PCR; detection

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