摘要
目的分析新型冠状病毒无症状感染者咽拭子样本中新冠病毒载量随时间的变化趋势,为新型冠状病毒防控提供科学依据。方法使用数字聚合酶链反应(Digital PCR,dPCR)和实时荧光定量聚合酶链反应(Quantitative realtime PCR,qPCR)技术对2例新型冠状病毒无症状感染者不同时间采集的咽拭子样本进行新冠核酸检测,同时使用胶体金法对采集的咽拭子样本进行新冠抗原检测。使用SPSS 20.0软件进行统计分析,配对卡方检验比较dPCR法与qPCR法、dPCR法与胶体金法以及10种胶体金试剂检测的阳性率差异,以P<0.05为差异有统计学意义。用Kappa系数判断结果一致性。结果新型冠状病毒无症状感染者A连续采集21份咽拭子样本,dPCR法检测ORF1ab基因的检测值为0~46080000 copies/ml,N基因检测值为0~45810000 copies/ml;新型冠状病毒无症状感染者B连续采集18份咽拭子样本,dPCR法检测ORF1ab基因检测值为0~138500000 copies/ml,N基因检测值为0~121600000 copies/ml。新型冠状病毒无症状感染者A21份咽拭子样本经dPCR法检出13份双靶基因阳性,1份单靶基因(ORF1ab)阳性,经qPCR法检出10份双靶基因阳性,胶体金法检出6份阳性。新型冠状病毒无症状感染者B18份咽拭子样本经dPCR法及qPCR法均检出15份双靶基因阳性,胶体金法检出5份阳性。dPCR法与qPCR法检测39份咽拭子样本中的ORF1ab基因及N基因,检测结果一致性为高度一致,且2种方法阳性率差异无统计学意义;胶体金法阳性率低于dPCR法,不同品牌的胶体金试剂检测的阳性率差异有统计学意义。结论2例新型冠状病毒无症状感染者咽拭子样本中核酸拷贝数随着采样时间延伸总体呈下降趋势,第15 d左右咽拭子样本中新冠核酸拷贝数趋于0;核酸检测法灵敏度高于胶体金法;需慎重选择胶体金试剂的品牌,以避免出现假阳性结果。
Objective To analyze the changing trend of COVID-19 viral load over time in throat swab samples from asymptomatic persons infected with COVID-19,and to provide scientific basis for the prevention and control of COVID-19.Methods Throat swabs were collected from two asymptomatic persons infected with COVID-19 at different times.Digital polymerase chain reaction(dPCR)and quantitative real-time PCR(qPCR)were used to detect COVID-19 nucleic acid in throat swab samples.COVID-19 antigen in throat swab samples was detected by colloidal gold method.SPSS 20.0 software was used for statistical analysis.The positive rates of dPCR and qPCR,d PCR and colloidal gold and 10 kinds of colloidal gold reagents were compared by paired Chi-square test.P<0.05 was considered to be statistically significant.Kappa coefficient was used to judge the consistency of results.Results A total of 21 throat swab samples of asymptomatic person A infected with COVID-19 were collected continuously.The detection value of ORF1ab gene by dPCR was 0-46080000 copies/ml,and the detection value of N gene was 0-45810000 copies/ml.A total of 18 throat swab samples were collected from asymptomatic person B infected with COVID-19.The detection value of ORF1ab gene by dPCR was 0-138500000 copies/ml,and the detection value of N gene was 0-121600000 copies/ml.In 21 throat swab samples of asymptomatic infected person A,13 dual-target genes and 1 single-target gene(ORF1ab)were detected positive by dPCR,10dual-target genes were detected positive by qPCR,and 6 samples were detected positive by colloidal gold method.In 18 throat swab samples of asymptomatic infected person B,15 dual-target genes were detected positive by dPCR and qPCR,and 5 samples were positive by colloidal gold method.Both d PCR and qPCR were used to detect ORF1ab gene and N gene in 39 throat swab samples.The results were highly consistent,and there was no statistical significance in the positive rate between the two methods.The positive rate of colloidal gold method was lower than that of dPCR metho
作者
廖虹瑜
冯玉亮
刘家洁
黄伟峰
周良君
LIAO Hongyu;FENG Yuliang;LIU Jiajie;HUANG Weifeng;ZHOU Liangjun(Sichuan Center for Disease Control and Prevention,Chengdu 610041,Sichuan Province,China;Mianyang Center for Disease Control and Prevention,Mianyang 621000,Sichuan Province,China)
出处
《寄生虫病与感染性疾病》
CAS
2023年第4期221-229,共9页
Parasitoses and Infectious Diseases
基金
四川省科技计划项目(项目编号:2021YFS0405)。
