现场PCR诊断箱POCKIT试剂联合应用技术快速检测炭疽芽孢杆菌的实验研究  

Experimental Study on Rapid detection of Bacillus Anthracis by PCR Diagnostic Kit Combined with POCKIT Technology

作　　者：尹世辉[1] 唐磊[1] 孙建飞 代伟萍[1] 袁爽[1] 邢智锋[1] 李杉杉[2] YIN Shihui;TANG Lei;SUN Jianfei;DAI Weiping;YUAN Shuang;XING Zhifeng;LI Shanshan(Heilongjiang center for disease control and prevention,Harbin,Heilongjiang 150030,China;The First Affiliated Hospital of Heilongjiang University of Traditional Chinese Medicine,Harbin,Heilongjiang 150030,China)

机构地区：[1]黑龙江省疾病预防控制中心,黑龙江哈尔滨150030 [2]黑龙江中医药大学附属第一医院,黑龙江哈尔滨150030

出　　处：《大医生》2019年第8期71-72,共2页Doctor

摘　　要：目的建立一种灵敏度高、特异性强、操作简单、检测结果准确并对检测环境要求低等优点的炭疽芽孢杆菌基因检测方法。方法利用Taqman探针原理,设计炭疽杆菌质粒PXO1和PXO2基因引物,采用冷冻干燥工艺将PCR检测试剂全部组分制备成冻干粉,使用时加入缓冲液进行溶解,与PCR诊断箱中配置便携PCR联合使用,可实现现场快速检测的目的。结果使用Taqman探针原理的PCR诊断箱和POCKIT试剂组合应用是目前最适合的现场检测方法,在敏感性和准确性方面一致,且操作简便。结论本研究建立的现场PCR诊断箱POCKIT试剂联合应用技术,方法可特异、灵敏地检测出患者的涂抹样本、环境样本。胶体金检测及荧光定量PCR检测结果充分证明此技术在现场快速检测炭疽疫情中具有重要意义。Objective To establish a method for the genetic detection of bacillus anthracis with high sensitivity,high specificity,simple operation,accurate detection results and low environmental requirements.Methods In accordance with that principle of Taqman probe,primers of Bacillus anthracis plasmid PXO1 and PXO2 gene were designed,and all components of PCR detection reagent were prepared into freeze-dried pow by freezedrying process,and buffer was added for dissolution in use.In combination with a portable PCR configure in a PCR diagnostic box,that purpose of on-site rapid detection can be achieved.Results The combination application of PCR diagnostic box and POCKIT reagent using Taqman probe principle is the most suitable field detection method at present,consistent in sensitivity and accuracy,and easy to operate.Conclusion The field PCR kit developed in this study can be used to detect smear samples and environmental samples in a specific and sensitive manner.The results of colloidal gold detection and fluorescence quantitative PCR can fully prove the significance of this technique in the field rapid detection of anthrax.

关 键 词：炭疽 PCR诊断箱 POCKIT试剂 质粒 现场检测 

分 类 号：S85[农业科学—兽医学]