含漱液中冠状病毒的即时检测研究(一)  

Study on detecting coronavirus in mouthwash fluid

作　　者：高星[1] 程钢[2] 何蕴绍[3] 林长樱[1] 刘海林[1] 贾明艳[4] 罗会英[1] 王如岗[1] 石维先[1] 丁立新[1] 韩丽丽[1] 黄芳[1] 

机构地区：[1]北京市疾病预防控制中心 [2]卫生部医药生物工程技术研究中心 [3]中山大学生物工程研究所 [4]北京市卫生局

出　　处：《中国公共卫生》2003年第5期F001-F003,共3页Chinese Journal of Public Health

基　　金：国家科技部;卫生部攻关指令性计划项目

摘　　要：目的　建立冠状病毒抗原早期快速诊断方法。方法　采集非典型肺炎病人 ( 16 3例 )、疑似病人 ( 2 8例 )、发热待查者 ( 17例 )、医学观察对象 ( 43人 )和正常人 ( 33人 )含漱液 ,应用实时荧光定量PCR(FQ -PCR)方法和试剂盒 ,进行冠状病毒抗原检测。结果　入院后 3d以内的非典型肺炎病例 93例 ,占 5 7.1%;入院第 4～ 2 0d 70例 ,占42 9%。 93例检出 5 3例冠状病毒抗原阳性者 ,总阳性率为 5 7 0 %;其中入院第 1d为 5 0 0 %,第 2d为 6 9 2 %,第 3d45 5 %。入院 4d后的 70例中检出 5名 ,阳性率仅为 7 1%。 2 8名疑似非典型肺炎患者含漱液阳性率为 3 4%;发热待查者、医学观察对象和正常人均未检出冠状病毒抗原阳性者。通过对含漱液中不同入院时间冠状病毒的检测 ,发现入院第 2d冠状病毒抗原阳性率最高 ,第 3d开始明显下降 ,第 4d以后已经达到很低的水平。结论　该检测方法度灵敏高 ,最低可检出 1个拷贝的基因 ;特异度高 ,能够满足临床诊断和医学观察对象和密切接触者筛查的要求 ;实时检测 ,准确定量 ,误差小 ;全封闭检测 ,杜绝产物污染导致的假阳性 ;单人份试剂包装 ,质量稳定 ,使用方便 ;符合卫生部和国家食品与药品监督管理局规定 ,可以大规模用于临床检验。Objective To search an instant early and rapid diagnostic method to determine coronavirus.Methods Firstly,mouthwash(NPA) fluid is collected from 163 SARS patients,28 suspected cases,43 medically observatory cases and 33 health individuals.FQ-PCR and reagent kits are used to test coronavirus.Results There are 93 (57.1%)SARS patients hospitalized within three days and 70 (42.9%)SARS patients hospitalized within four to twenty days.53 of the 93 patients are tested with positive coronavirus.The total positive rate is 57.0%.Furthermore,the positive rate in the first,second and third day of hospitalization are 50.0%,69.2% and 45.5% respectively.The total positive rate of the 70 patients hospitalized more than four days is only 7.1%.That is 3.4% among 28 suspected cases.The positive rate is negative among the 17 hospitalized cases with fever and waiting for determination,43 medically observatory cases and 33 health individuals.Through testing coronavirus in mouthwash fluid in different incidence time,it can be found that virus positive rate of the coronavirus is highest in the second day of hospitalization.The positive rate begins to decline in the third day of hospitalization and reaches the lowest level after the fourth day of hospitalization.Conclusion The method has high sensitivity,specificity and can detect at least one copy of gene.It can meet the demands for clinical diagnosis as well as screening and detecting medically observatory cases and closely contacting persons.It can be used at all times to determine the quantities exactly with little error.The method is used under full-sealed conditions,which can eliminate false positive result.The package is for one person with stable quality and facility in use.It satisfies the demands of Ministry of Public Health and Supervisory of National Food and Drug Administrative Bureau.It can be used in clinical diagnosis extensively.

关 键 词：冠状病毒 含漱液 实时荧光定量PCR法 

分 类 号：R373.1[医药卫生—病原生物学] Q782[医药卫生—基础医学]