The Detection Results of Viral Nucleic Acid in Various Biological Samples of Patients with COVID-19 in Kunming
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摘要:
目的 通过国产2019-nCoV(新型冠状病毒)核酸测定试剂,测定确诊的COVID-19(新型冠状病毒肺炎)患者的多种生物样本内SARS-CoV-2 核酸的携病毒情况,对机体不同系统中SARS-CoV-2的状态展开观察,并分析其临床价值,为诊断和治疗提供参考。 方法 收集昆明市第三人民医院53例确诊的新型冠状病毒肺(COVID-19)患者的鼻拭子、咽拭子、痰液、粪便和尿液样本,实时荧光逆转录聚合酶链反应(RT-PCR)测定法,来平行测定病毒ORF1ab基因与N基因核酸,比较不同类型样本的阳性率和一致性。 结果 患者的多种生物标本核酸Ct值的变化与病程都高度相关,发病15 d前后来分析核酸检测结果,发现在发病15 d后病毒核酸Ct值与15 d内相比病毒核酸载量,差异有统计学意义(P < 0.05)。在发病第1~7天和8~14 d,鼻拭子阳性检出率为88.67%(47/53)和79.24%(42/53),而咽拭子阳性检出率为73.58%(39/53)和53.85%(28/52)。粪便样本核酸检测阳性率12.28%(7/57),尿液样本均未检测出病毒核酸。 结论 在新冠肺炎患者发病的不同阶段,鼻拭子、咽拭子、痰液样本有较高的核酸检出率;在阳性检出率方面,鼻拭子相较咽拭子偏高;SARS-CoV-2有粪-口传播可能,同时病毒液可能会潜在感染消化道。 -
关键词:
- 2019新型冠状病毒 /
- 新型冠状病毒肺炎 /
- 实时荧光逆转录聚合酶链反应 /
- 核酸
Abstract:Objective To detect the viral status of SARS-CoV-2 nucleic acid in various biological samples of patients diagnosed with New Coronavirus pneumonia (COVID-19) by using the nucleic acid reagents of New Coronavirus (2019-nCoV). The status and clinical significance of SARS-CoV-2 in different body systems were observed, providing reference for diagnosis and treatment. Methods The nasal swabs, throat swabs, sputum, feces and urine samples of 53 patients diagnosed with New Coronavirus lung (COVID-19) in Kunming Third People's Hospital were collected. Real-time fluorescence reverse transcription polymerase chain reaction (RT-PCR) was used to detect the ORF1ab gene and N gene nucleic acid in parallel. The positive rates and consistency of different samples were compared. Results The results showed that the changes of CT value of nasal swab, pharyngeal swab, sputum sample and fecal sample were highly correlated with the course of disease. The analysis of nucleic acid detection results 15 days before and after the onset showed that the difference of CT value of viral nucleic acid 15 days after the onset was statistically significant compared with that within 15 days (P < 0.05). The positive rate of nasal swab was 88.67% (47/53) and 79.24% (42/53), while that of pharyngeal swab was 73.58% (39/53) and 53.85% (28/52). The positive rate of nucleic acid detection in stool samples was 12.28% (7/57), and no viral nucleic acid was detected in urine samples. Conclusion In novel coronavirus pneumonia patients at different stages of onset, nasal swabs, throat swabs and sputum samples show higher nucleic acid detection rates. The positive rate of nasal swab is higher than that of pharyngeal swab. SARS-CoV-2 has the possibility of fecal oral transmission, and the digestive tract may be the potential target organ of virus infection. -
表 1 不同病程不同生物样本新型冠状病毒核酸检测
Table 1. Detection of SARS-CoV-2 nucleic acid in different biological samples with different course of diseas
收集日期 样本类型 新型冠状病毒肺炎病例 (COVID-19 ) χ2 P 例数 n 阳性数 阳性率 (%) 0~7 d 鼻拭子 53 47 88.67 15.55 < 0.001* 阳性 咽拭子 53 39 73.58 痰液 50 27 54.00 粪便 / / / 尿液 / / / 8~14 d 鼻拭子 53 42 79.24 88.08 < 0.001* 阳性 咽拭子 52 28 53.85 痰液 49 16 32.65 粪便 38 5 13.16 尿液 53 0 0.00 ≥15 d 鼻拭子 43 6 13.95 5.78 0.216 阳性 咽拭子 45 2 4.44 痰液 27 3 11.11 粪便 19 2 10.52 尿液 28 0 0.00 *P < 0.05。 -
[1] Du R H,Liang L R,Yang C Q,et al. Predictors of mortality for pa- tients with COVID-19 pneumonia caused by SARS-CoV-2:aprospective cohort study[J]. Eur Respir J,2020,55(5):2000524. doi: 10.1183/13993003.00524-2020 [2] Jin X,Lian J S,Hu J H,et al. Epidemiological,clinical and virological characteristics of 74 cases of coronavirus-infected disease 2019 (COVID-19) with gastrointestinal symptoms[J]. Gut,2020,69(6):1002-1009. doi: 10.1136/gutjnl-2020-320926 [3] Zhou F,Yu T,Du R,et al. Clinical course and risk factors for mortality of adult inpatients with COVID-19 in Wuha,China:A retrospective cohort study[J]. Lancet,2020,395(10229):1054-1062. doi: 10.1016/S0140-6736(20)30566-3 [4] WHO coronavirus disease(COVID-19) dashboard[Z]. 2020. https://covid19.who.int [5] 国家卫生健康委. 新型冠状病毒感染的肺炎诊疗方案(试行第八版 修订版)国家卫生健康委(2020-4-14)[EB/OL] .http://www.nhc.gov.cn/xcs/zhengcwj/202008/0a7bdf12bd4b46e5bd28ca7f9a7f5e5a/files/a449a3e2e2c94d9a856d5faea2ff0f94.pdf [6] 中华人民共和国国家卫生健康委员会 . 《新型冠状病毒感染的肺炎诊疗方 案(试行第七版)》(2020-03-03)[EB/OL]. http://www. nhc. gov. cn/yzygj/s7653p/202003/46c9294a7dfe4cef80dc7f5912eb1989.shtml. [7] 国家卫生部办公厅, 国家卫生健康委员会医政医管局办公室. 医疗机构临床基因扩增检验实验室管理办法[EB/OL]. (2010-12-06). https://www.51mch.com/uploadfiles/2016/11/201611151432113211.pdf [8] 国家卫生健康委办公厅, 科技教育司办公室. 新型冠状病毒实验室生物安全指南(第二版)[OB/OL]. (2020-01-23). http://news.bioon.com/article/6749573.html [9] 国家卫生健康委办公厅, 国家科技教育司办. 关于印发新型冠状病毒实验室生物安全指南(第二版)的通[OB/OL]. (2020-01-23 ). https://news.sciencenet.cn/htmlnews/2020/1/435132.shtm [10] Bogoch I I,Watts A,Thomas-Bachli A,et al. Potential for global spread of a novel coronavirus from China[J]. J Travel Med,2020,27(2):taaa011. doi: 10.1093/jtm/taaa011 [11] Lu R,Zhao X,Li J,et al. Genomic characterisation and epidemiology of 2019 novel coronavirus:Implications for virus origins and receptor binding[J]. Lancet,2020,395(10224):565-574. doi: 10.1016/S0140-6736(20)30251-8 [12] Su S,Wong G,Shi W,et al. Epidemiology,genetic recombination,and pathogenesis of coronaviruses[J]. Trends Microbiol,2016,24(6):490-502. doi: 10.1016/j.tim.2016.03.003 [13] Zhu N,Zhang D,Wang W,et al. A novel coronavirus from patients with pneumonia in China,2019[J]. N Engl J Med,2020,382(8):727-733. doi: 10.1056/NEJMoa2001017 [14] 施绍瑞,聂滨,郭渝,等. 新型冠状病毒肺炎病例多种生物样本的病毒核酸检测结果[J]. 华西医学,2020,35(2):132‐136. [15] 刘焱斌,刘涛,崔跃,等. 鼻拭子与咽拭子两种取样方法在新型冠状病毒肺炎核酸筛检中的比较研究[J]. 中国呼吸与危重监护杂志,2020,19(2):141-143. [16] Zou L R,Ruan F,Huang M X,et al. SARS-CoV-2 viral load in upper respiratory specimens of infected patients[J]. N Engl J Med,2020,382(12):1177-1179. doi: 10.1056/NEJMc2001737 [17] XU Z,SHI L,WANG Y J,et al. Pathological findings of COVID-19 associated with acute respiratory distress syndrome[J]. Lancet Respir Med,2020,8(4):420-422. doi: 10.1016/S2213-2600(20)30076-X [18] Zhang H, Kang Z, Gong H, et al. The digestive system is a potential route of 2019- nCov infection: a bioinformatics analysis based on single-cell transcriptomes [J/OL]. https://doi. org/10.1101/ 2020.01.30.927806. [19] 俞龙,彭伟,朱丹. 2019 冠状病毒病患者出院前核酸检 测粪便标本阳性而肛拭、咽拭子阴性1例报告[J]. 解放军医学院学报,2020,41(2):125-126. doi: 10.3969/j.issn.2095-5227.2020.02.008 [20] 吴建国,罗建飞,刘家盛,等. 重型/危重型新型冠状病毒肺炎患者多种生物样本核酸检测结果分析[J]. 解放军医学院学报,2020,41(3):205-207,211. doi: 10.3969/j.issn.2095-5227.2020.03.001 [21] 魏秋华,任哲. 2019新型冠状病毒感染的肺炎疫源地消毒措施[J]. 中国消毒学杂志,2020,37(1):59-62. doi: 10.11726/j.issn.1001-7658.2020.01.019