PMA-PCR诊断技术研究进展  

Advances in the Researches on PMA-PCR Technology

作　　者：石艳萍 陈弟诗[1] 王印[2] 陈斌[1] 杨悦 张鹏飞[2] 李春[1] 邵靓[1] Shi Yanping;Chen Dishi;Wang Yin;Chen Bin;Yang Yue;Zhang Pengfei;Li Chun;Shao Jing(Sichuan Center for Animal Disease Prevention and Control,Chengdu,Sichuan 610041,China;Sichuan Agricultural University,Chengdu,Sichuan 611130,China)

机构地区：[1]四川省动物疫病预防控制中心,四川成都610041 [2]四川农业大学,四川成都611130

出　　处：《中国动物检疫》2023年第3期72-78,共7页China Animal Health Inspection

基　　金：四川省科技厅应用基础研究项目(2021YJ0270);四川省科技厅重点研发项目(2020YFN0034)。

摘　　要：叠氮溴化丙锭(propidium monoazide,PMA)是一种光反应的DNA结合染料,具有极高的DNA亲和结合能力。PMA可进入死亡和外膜损伤严重的微生物内部,插入DNA结构中,经可见光刺激发生光裂解后,同DNA结合发生共价交联反应,共价交联产物可以抑制DNA的PCR扩增。基于PMA的这一特性,PMAPCR可用于鉴别活死细菌、真菌、病毒等微生物。在实际应用中,PMA-PCR检测结果受样本浊度、活死微生物比例、PMA浓度、暗处理条件(温度、暗处理时间)、曝光条件(光源、光处理时间)、样本处理方法以及目的基因扩增长度等多种因素影响。PMA-PCR已被应用于动植物疫病防控、食品安全、公共卫生等多个领域,具有广阔的应用前景。Propidium monoazide(PMA)was a kind of photoreactive DNA-binding dye with extremely high affinity for DNA.PMA could enter into the dead or membrane-damaged microorganisms,then insert into DNA structure,and following photolysis stimulated by visible light,bind to DNA to produce covalent crosslinking reaction.PCR amplification of DNA could be inhibited by the products.Based on the above characteristics,PMA-PCR technology could be used to identify living bacteria,fungi,viruses and other microorganisms.In practice,the detection results by PMA-PCR could be affected by sample turbidity,the ratio of dead and living microorganisms,PMA concentration,conditions of dark processing(temperature,dark processing time),exposure conditions(light source,light processing time),treatment method for samples,the amplification length of target genes and other factors.PMA-PCR,with wide prospects of application,had been used in animal and plant disease prevention and control,foodsafety,public health and other areas.

关 键 词：叠氮溴化丙锭 PMA PCR 检测 

分 类 号：S852.23[农业科学—基础兽医学]