畜牧与兽医2018，Vol.50Issue(5)：89-92,4.
斑点叉尾鮰病毒焦磷酸测序检测方法的建立
Establishment of pyrosequencing assay for detection of channel catfish virus
摘要
Abstract
The aim of the study was to establish a pyrosequencing assay for rapid detection and identification of channel catfish virus (CCV).The amplification primers and sequencing primer were designed for the conserved region of protein kinase gene of CCV.After PCR reaction,the product was detected with pyrosequencing and identified by genomic alignment.The pyrosequencing assay was established of the PK gene for CCV by optimizing the reaction conditions and system.The results showed that the PCR product was 144bp and it could specifically identify the target viruses.The minimum detection limit of the nuclei acid was 5× 10-6ng/μL Pyrosequencing repeated three times accurately determined the nucleotide sequence to be 45bp long.It was demonstrated that the pyrosequencing assay established here possessed good specificity,high sensitivity and stability in detection of CCV,The assay is applicable to r high throughput and time-consuming detection of CCV.It takes only 4 h.关键词
斑点叉尾鮰病毒/焦磷酸测序/PCR/蛋白激酶基因/检测Key words
channel catfish virus/pyrosequencing/PCR/protein kinase gene/detection分类
农业科技引用本文复制引用
王娜,张旻,景宏丽,江育林,吴绍强..斑点叉尾鮰病毒焦磷酸测序检测方法的建立[J].畜牧与兽医,2018,50(5):89-92,4.基金项目
国家“十二·五”科技支撑计划(2013BAD12B02) （2013BAD12B02）
