中国兽医科学2024，Vol.54Issue(3)：325-329,5.DOI:10.16656/j.issn.1673-4696.2024.0041
猪库布病毒纳米PCR快速检测方法的建立
Establishment and preliminary application of nano-PCR method for detecting porcine Kubuvirus
摘要
Abstract
In order to establish a method for rapid diagnosis of porcine kobuvirus and provide a more accurate and convenient method for clinical detection of the virus,this experiment designed a pair of specific primers to amplify the VPO region of the PKV genome.Nano-gold particle materials were added to the traditional PCR reaction system,and after optimization,a convenient,fast and specific nano-PCR method for detecting PKV was established.The test results show that this method can specifi-cally amplify porcine Kubuvirus and has no cross-reactivity with TGEV,PEDV,PDCoV,PRV and PSaV.The nano-PCR showed a higher sensitivity compared to the normal PCR,with a 1 000-fold increased sensitivi-ty.The lowest concentration of nucleic acid detection for ordinary PCR was 5.61 × 10-7 ng/μL,while the lowest nucleic acid detection concentration for nano-PCR was only 5.61 × 10-10 ng/μL.Thirty feces and blood clinical samples from pigs with diarrhea from different pig farms were tested.The results showed that the positive detection rate of nano-PCR was 23.33％(7/30).The positive detection rate of ordinary PCR is 16.66％(5/30).Therefore,the nano-PCR method established in this trial showed higher specifici-ty and sensitivity in the detection of porcine Kubuvirus,which has important clinical significance for the rapid diagnosis and detection of porcine Kubuvirus.关键词
猪库布病毒/nano-PCR/纳米金颗粒Key words
porcine kobuvirus/nano-PCR/gold nanoparticle分类
农业科技引用本文复制引用
李睿,凤焱,孙亮,姜秋杰,呼延含荣,张秀森,程敏,郭利,赵立峰..猪库布病毒纳米PCR快速检测方法的建立[J].中国兽医科学,2024,54(3):325-329,5.基金项目
吉林省科技厅科技发展计划项目(YDZJ202203CGZH050,20210202101NC) （YDZJ202203CGZH050,20210202101NC）
吉林省农业关键核心技术示范推广项目(202300702) （202300702）
