含漱液中冠状病毒的即时检测研究(一)

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(163例)、疑似病人(28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57．1％；入院第4—20d70例，占42．9％。93例检出53例冠状病毒抗原阳性考，总阳性率为57．0％；其中入院第ld为50．0％，第2d为69．2％，第3d45．5％。入院4d后的70例中检出5名，阳性率仅为7．1％。28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末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。

Study on detecting coronavirus in mouthwash fluid

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.