Establishment and Application of PCR Method for Detecting Psittacine Beak Feather Virus
In order to establish a sensitive and specific PCR method for detecting psittacosis virus,and to provide technical support for rapid clinical diagnosis of psittacosis.According to the gene sequence of Psittacosis virus published in Genbank,the highly conserved region was identified by comparative analysis,a pair of detection primers was designed to extract the DNA template of psittacosis virus positive samples,and a series of adjustments were made to the amplification system and reaction conditions to determine the ideal PCR reaction scheme.The sensitivity limit of the method was determined by the 10-fold gradient dilution DNA template method.The method was used to detect infectious bursitis virus,avian infectious bronchitis virus,Newcastle disease virus,infectious laryngotracheitis virus,fowlpox virus and Group I avian adenovirus type 4,and to determine the specificity of the method.The PCR method was used to detect 90 clinical samples and test its practicability.The results showed that the designed detection primer could specifically amplify PBFDV,and the other 6 poultry viruses were negative.The lower limit of DNA concentration for detection was 3.725×10-6 ng/µL.Clinical application showed that 28 of 90 suspected samples were positive,with a positive rate of 31.1%.The results showed that a sensitive and specific PCR method for psittacosis virus was established successfully.
国家科技期刊平台
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