Abstract:The outbreak of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)all over the world leads to serious respiratory diseases and high mortality,which arouses wide concern. No specific treatment for coronavirus disease 2019(COVID-19)caused by the virus has been found,so early diagnosis is the main prevention and control measure at present. According to the COVID-19 Diagnosis and Treatment Plan(Seventh Trial),Being positive in nucleic acid testing or in antibody testing,or being highly homologous in gene sequencing can confirm SARS-CoV-2 infection. However, all these detection Methods have potential false negative and false positive Results. This article summarizes the SARS-CoV-2 laboratory testing Methods that are being applied and studied at present,such as virus isolation,second generation sequencing,reverse transcription-polymerase chain reaction,nucleic acid isothermal amplification,gene chip and antibody detection,and discusses their advantages and disadvantages to optimize the diagnosis of SARS-CoV-2 infection.
[1] 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. [2] GORBALENYA A E,BAKER S C, BARIC R S, et al. Severe acute respiratory syndrome-related coronavirus:the species and its viruses-a statement of the Coronavirus Study Group [J/OL]. BioRxiv(2020-02-07)[2020-03-17].https://doi.org/10.1101/ 2020.02.07.937862. [3] TO K K, TSANG O T,CHIK-YAN YIP C,et al. Consistent detection of2019 novel coronavirus in Saliva [J/OL]. Clin Infect Dis(2020-02-12)[2020-03-17]. https://academic.oup.com/cid/article-lookup/doi/10.1093/cid/ciaa149. [4] HANG W,DU R H, LI B,et al.Molecular and serological investigation of 2019-nCoV infected patients: implication of multiple shedding routes[J]. Emerg Microbes Infect, 2020, 9(1): 386-389. [5] XIA J, TONG J,LIU M,et al. Evaluation of coronavirus in tears and conjunctival secretions of patients with SARS-CoV-2 infection [J]. J Med Virol(2020-02-26)[2020-03-17]. https://doi.org/10.1002/jmv.25725. [6] 邓少丽,刘丁.新型冠状病毒肺炎的实验室分子诊断及生物安全刍识[J/OL].国际检验医学杂志[2020-03-17].http://kns.cnki.net/kcms/detail/50.1176.R.20200310.0935.002.html. [7] 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核酸筛检中的比较研究 [J/OL] 中国呼吸与危重监护杂志,2020(2020-03-02)[2020-03-17].http://kns.cnki.net/kcms/detail/51.1631.R.20200228.1506.004.html. [8] YANG Y,YANG M H, SHEN C G,et al. Evaluating the accuracy of different respiratory specimens in the laboratory diagnosis and monitoring the viral shedding of2019-nCoV infections[J]. medRxiv[2020-03-17].https://doi.org/10.1101/2020.02.11.20021493. [9] 中华人民共和国国家卫生健康委员会办公厅.新型冠状病毒感染的肺炎实验室检测技术指南(第四版)[EB/OL].(2020-02-06) [2020-03-17].http://www.nhc.gov.n/jkj/s3577/202002/573340613cab 243b3a7f61df260551dd4.shtml. [10] 张欢,郑焕英,邹丽容,等. 广东省首例新型冠状病毒的分离与鉴定 [J/OL]. 病毒学报,2020(2020-03-05 )[2020-03-17]https://doi.org/10.13242/j.cnki.bingduxuebao.003657. [11] 许金和, 王水良, 张胜行,等. 新型冠状病毒核酸检测方法 [J/OL]. 国际检验医学杂志, 2020(2020-03-03)[2020-03-17].http://kns.cnki.net/kcms/detail/50.1176.R.20200303.1428.002.html. [12] 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. [13] WANG M, FU A, HU B,et al. Nanopore target sequencing for accurate and comprehensive detection of SARS-CoV-2 and other respiratory viruses[EB/OL].(2020-03-04)[2020-03-17].https://doi.org/10.1101/2020.03.04.20029538. [14] 鲁彦, 居军, 李德红. 核酸和血清学指标结合,多类型联检,提高新型冠状病毒检出率 [J/OL]. 检验医学与临床, 2020(2020-03-10)[2020-03-17].http://www.nhc.gov.cn/jkj/s3577/202002/573340613ab243b3a7f61df260551dd4/files/c791e5a7ea5149f680fdcb34dac0f54e.pdf. [15] 郭晓波,蔺京,来春艳,等. 新型冠状病毒与实时荧光RT-PCR核酸检测[J]. 陕西医学杂志,2020,49(3):264-266. [16] 梁海燕,刘文鑫,杨志刚. 等温核酸扩增技术进展[J]. 中国医学创新,2017,14(16):145-148. [17] CHEN C M, OUYANG S, LIN L Y, et al. Diagnostic accuracy of LAMP assay for HBV infection [J/OL]. J Clin Lab Anal(2020-03-10)[2020-03-17].https://doi.org/10.1002/jcla.23281. [18] HUANG P, WANG H, CAO Z, et al.A rapid and specific assay for the detection of MERS-CoV[J]. Front Microbiol, 2018, 9:1101-1109. [19] KEIGHTLEY M C, SILLEKENS P, SCHIPPERS W, et al.Real-time NASBA detection of SARS-associated coronavirus and comparison with real-time reverse transcription-PCR[J]. J Med Virol, 2005, 77(4): 602-608. [20] 高闪电, 常惠芸, 丛国正,等. NASBA(依赖核酸序列的扩增)技术及其在病毒检测中的应用[J]. 中国生物工程杂志, 2009,29(1): 80-85. [21] WANG B, POTTER S J, LIN Y, et al.Rapid and sensitive detection of severe acute respiratory syndrome coronavirus by rolling circle amplification[J]. J Clin Microbiol,2005, 43(5): 2339-2344. [22] WANG D, URISMAN A, LIU Y T, et al.Viral discovery and sequence recovery using DNA microarrays[J]. PLoS Biol, 2003, 1(2): E2. [23] CaptitalBio Corporation.CapitalBio Respiratory Virus Nucleic Acid Detection Kit[EB/OL].[2020-03-17]. http://www.capitalbio.com/diagnostic/tbrelatedkits/identificationarray/RePaKit/28042.shtml. [24] JUANG J L, CHEN T C, JIANG S S, et al.Coupling multiplex RT-PCR to a gene chip assay for sensitive and semiquantitative detection of severe acute respiratory syndrome-coronavirus[J]. Lab Invest, 2004, 84(9): 1085-1091. [25] GUO X, GENG P, WANG Q, et al.Development of a single nucleotide polymorphism DNA microarray for the detection and genotyping of the SARS coronavirus[J]. J Microbiol Biotechnol, 2014, 24(10): 1445-1454. [26] 罗效梅, 王静, 张娅,等. 全血SARS-CoV-2特异性抗体检测对2019-冠状病毒病的临床应用价值分析 [J/OL]. 西南大学学报(自然科学版)(2020-09-09)[2020-03-17].http://kns.cnki.net/kcms/detail/50.1189.N.20200306.1716.004.html. [27] 中华人民共和国国家卫生健康委员会办公厅,中华人民共和国国家中医药管理局办公室.关于印发新型冠状病毒肺炎诊疗方案(试行第七版)的通知[EB/OL].(2020-03-04)[2020-03-17].http://www.nhc.gov.cn/yzygj/s7653p/202003/46c9294a7dfe4cef80dc7f5912eb1989.shtml. [28] LI Z, YI Y, LUO X, et al. Development and clinical application of arapid IgM-IgG combined antibody test for SARS-CoV-2 infection diagnosis [J/OL]. J Med Virol, 2020(2020-02-27)[2020-03-17].https://onlinelibrary.wiley.com/doi/abs/10.1002/jmv.25727. [29] 马建, 贺鹏飞, 赵晨燕,等. SARS和MERS中和抗体假病毒检测方法的建立[J]. 病毒学报, 2019, 35(2): 189-195. [30] 薛雄燕, 朱嫦琳, 黄少珍,等. 灭活血液样本对不同方法检测2019新型冠状病毒抗体检测结果的影响 [J/OL]. 南方医科大学学报, 2020(2020-03-02)[2020-03-17].http://kns.cnki.net/kcms/detail/44.1627.R.20200228.1733.002.html. [31] 邹明园,吴国球. 抗原交叉反应对新型冠状病毒血清特异性抗体检测的影响[J]. 临床检验杂志, 2020: 1-3. [32] CHE X Y, HAO W, WANG Y, et al.Nucleocapsid Protein as Early Diagnostic Marker for SARS[J]. Emerg Infect Dis, 2004, 10(11):1947-1949. [33] CHEN Y, CHAN K H, KANG Y, et al.A sensitive and specific antigen detection assay for Middle East respiratory syndrome coronavirus[J]. Emerg Microbes Infect, 2015, 4(4): e26. [34] 鲍春梅,李波, 王欢, 等. 新型冠状病毒的实验室生物风险评估与风险控制[J]. 传染病信息, 2020, 33(1): 90-95.