关闭
 
读者在线:用户名 密码
首页 期刊简介 投稿须知 期刊目录 专家风采 编委会 特邀顾问 联系我们 移动出版
  1. 1
  2. 2
  3. 3
  4. 4
  5. 5



刊物信息

期刊名称:药物分析杂志
主管单位:中国科学技术协会
主办单位:中国药学会
承办:中国食品药品检定研究院
主编:金少鸿
地址:北京天坛西里2号
邮政编码:100050
电话:010-67012819,67058427
电子邮箱:ywfx@nicpbp.org.cn
国际标准刊号:ISSN 0254-1793
国内统一刊号:CN 11-2224/R
邮发代号:2-237
 

访问统计
您是第  4 8 5 1 4 0 4 位浏览者
您当前的位置:首页 >> 正文

S/D残留量对SARS-CoV-2假病毒中和抗体检测方法的影响

Influence of S/D residue on SARS-CoV-2 neutralizing antibody evaluation

分类号:R917
出版年·卷·期(页码):2020,40 (4):613-619
DOI: 10.16155/j.0254-1793.2017.01.01
-----摘要:-------------------------------------------------------------------------------------------

目的: 评价有机溶剂/表面活性剂(solvent and detergent,S/D)法灭活后S/D残留量对严重急性呼吸综合征冠状病毒2(SARS-CoV-2)的假病毒中和抗体检测方法(pseudotyped virus based neutralization assay,PBNA)的影响。方法: 在严重急性呼吸综合征(severe acute respiratory syndrome,SARS)和2019冠状病毒病(coronavirus disease,COVID-19)患者恢复期血浆中加入0.1~10倍《中华人民共和国药典》(简称《中国药典》)规定残留标准[100 μg·mL-1聚山梨酯80(吐温80)+10 μg·mL-1磷酸三丁酯(TNBP)]浓度的S/D,模拟灭活后S/D残留量。用PBNA检测加入S/D前后的中和抗体水平,比较S/D对中和方法的影响。将不同浓度的S/D加入到SARS-CoV和SARS-CoV-2假病毒中进行灭活处理1、2和6 h,检测假病毒的滴度,评价S/D对冠状病毒假病毒的灭活效果。结果: 当S/D残留低于《中国药典》残留上限(100 μg·mL-1吐温80+10 μg·mL-1 TNBP)时,SARS-CoV-2中和抗体EC50平均值与无S/D组的PBNA检测结果无统计学差异;当S/D残留大于2.5倍《中国药典》残留标准时,S/D对PBNA有显著影响。含有10 mg·mL-1吐温80+1 mg·mL-1 TNBP的S/D试剂,室温静置1 h即可以灭活SARS-CoV和SARS-CoV-2假病毒,假病毒滴度下降分别达到5.11和4.80 log滴度,其灭活效果随着S/D的浓度下降而下降,S/D浓度为1%时无灭活效果。结论: S/D残留量对SARS-CoV-2的PBNA有显著影响,为保障PBNA的检测准确性,血浆中S/D残留量应不超过《中国药典》规定的标准,S/D试剂具有灭活2种冠状病毒假病毒的作用。

-----英文摘要:---------------------------------------------------------------------------------------

Objective: To evaluate the influence of residual solvent and detergent(S/D),which is used to inactivate SARS-CoV-2 in convalescent plasma(CP),on pseudotyped virus based neutralization assay(PBNA). Methods: To simulate S/D residue after inactivation,S/D from 0.1 to 10-fold amounts of what is listed in Pharmacopoeia of the People's Republic of China (Chinese Pharmacopoeia)[100 μg·mL-1 polysorbate 80(Tween 80) + 10 μg·mL-1 tributyl phosphate(TNBP)] were added into CPs from SARS(severe acute respiratory syndrome,SARS) or COVID-19 (coronavirus disease,COVID-19) infected individuals. The titers of neutralizing antibody before and after S/D addition were detected by PBNA. The influence of S/D on the detection was investigated. After 1,2 and 6 hours of treatment,the titers of pseudotyped virus were detected to evaluate the inactivation effect of S/D on coronavirus pseudotyped viruses. Results: When S/D residue did not exceed the residue standard (100 μg·mL-1 Tween 80+10 μg·mL-1 TNBP) stipulated in Chinese Pharmacopoeia,no significant difference was observed between the S/D and control groups for EC50 values of SARS-CoV-2 neutralizing antibody. When S/D residue exceeded the 2.5 folds of residue standard,a significant inhibitory effect was presented on neutralizing antibody detection. S/D containing 10 mg·mL-1 Tween 80+1 mg·mL-1 TNBP,could inactivate both pseudotyped viruses after 1-hour incubation at room temperature. The titers of pseudotyped virus decreased 5.11 and 4.80 log for SARS-CoV and SARS-CoV-2 respectively. When the S/D concentration was reduced to 1%,the inactivation effect disappeared. Conclusion: Residual S/D has significant inhibitory effect on evaluating neutralizing antibody when its concentration is higher than its standard in Chinese Pharmacopoeia. To ensure the accuracy of PBNA detection,the S/D residue in plasma should not exceed the standard in Chinese Pharmacopoeia. S/D reagent has the function of inactivating SARS-CoV and SARS-CoV-2 pseudotyped viruses.

-----参考文献:---------------------------------------------------------------------------------------
[1] REN LL,WANG YM,WU ZQ,et al.Identification of a novel coronavirus causing severe pneumonia in human:a descriptive study[J].Chin Med J(Engl),2020 Feb 11.doi:10.1097/CM9.0000000000000722
[2] 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
[3] WU F,ZHAO S,YU B,et al.A new coronavirus associated with human respiratory disease in China[J].Nature,2020,579(7798):265
[4] ZHOU P,YANG X L,WANG X G,et al.A pneumonia outbreak associated with a new coronavirus of probable bat origin[J].Nature,2020,579(7798):270
[5] WHO Director-General's Remarks at the Media Briefing on 2019-nCoV on 11 February 2020[EB/OL].[2020-02-11].https://www.who.int/dg/speeches/detail/who-director-general-s-remarks-at-the-media-briefing-on-2019-ncov-on-11-february-2020.
[6] Coronaviridae Study Group of the International Committee on Taxonomy of Viruses.The species severe acute respiratory syndrome-related coronavirus:classifying 2019-nCoV and naming it SARS-CoV-2[J].Nat Microbiol,2020 Mar 2.doi:10.1038/s41564-020-0695-z.[Epub ahead of print]
[7] FLEXNER S,LEWIS PA.Experimental epidemic poliomyelitis in monkeys[J].J Exp Med,1910,12(2):227
[8] LEIDER JP,BRUNKER PA,NESS PM.Convalescent transfusion for pandemic influenza:preparing blood banks for a new plasma product?[J].Transfusion,2010,50(6):1384
[9] MUPAPA K,MASSAMBA M,KIBADI K,et al.Treatment of Ebola hemorrhagic fever with blood transfusions from convalescent patients.International Scientific and Technical Committee[J].J Infect Dis,1999,179(Suppl 1):S18
[10] CHENG Y,WONG R,SOO YO,et al.Use of convalescent plasma therapy in SARS patients in Hong Kong[J].Eur J Clin Microbiol Infect Dis,2005,24(1):44
[11] ARABI Y,BALKHY H,HAJEER AH,et al.Feasibility,safety,clinical,and laboratory effects of convalescent plasma therapy for patients with Middle East respiratory syndrome coronavirus infection:a study protocol[J].Springerplus,2015,4:709
[12] LAN L,XU D,YE G,et al.Positive RT-PCR test results in patients recovered from COVID-19[J].JAMA,2020 Feb 27.doi:10.1001/jama.2020.2783.[Epub ahead of print]
[13] RABENAU HF,BIESERT L,SCHMIDT T,et al.SARS-coronavirus(SARS-CoV) and the safety of a solvent/detergent(S/D) treated immunoglobulin preparation[J].Biologicals,2005, 33(2):95
[14] 国家卫生健康委办公厅国家中医药管理局办公室.新型冠状病毒肺炎诊疗方案(试行第六版)[S] 2020 General Office of the national health and Health Commission,Office of State Administration of Traditional Chinese Medicine.Diagnosis and Treatment of Novel Coronavirus Pneumonia(6th trial version)[S].2020
[15] 中国研究型医院学会危重医学专业委员会,中国研究型医院学会危重医学专委会青年委员会.重型和危重型新型冠状病毒肺炎诊断和治疗专家共识[J].中华危重病急救医学,2020,32(2):129 Chinese Research Hospital Association of Critical Care Medicine,Youth Committee of Chinese Research Hospital Association of Critical Care Medicine.Chinese experts consensus on diagnosis and treatment of severe and critical new coronavirus pneumonia[J].Chin Crit Care Med,2020,32(2):129
[16] 杨晓明,侯继峰.康复期血浆应用于急性病毒性传染病现状及其治疗新型冠状病毒肺炎前景[J/OL].中国生物制品学杂志,2020[2020-03-12].https://doi.org/10.13200/j.cnki.cjb.002999 YANG XM,HOU JF.Current situation of novel coronavirus pneumonia in convalescent plasma and its prospect for treatment of acute viral infectious diseases[J/OL].Chin J Biol,2020[2020-03-12].https://doi.org/10.13200/j.cnki.cjb.002999
[17] NIE JH,LI QQ,WU JJ,et al.Establishment and validation of a pseudo-virus neutralization assay for SARS-CoV-2[J].Emerg Microbes Infect,2020,9(1):680
[18] LI Q,LIU Q,HUANG WJ,et al.Current status on the development of pseudoviruses for enveloped viruses[J].Rev Med Virol,2018,28:e1963
[19] 国家药品监督管理局药品审评中心.国药监注[2002] 160号血液制品去除灭活病毒技术方法及验证指导原则[S].2002:1 Drug Evaluation Center of the State Drug Administration.GJZ[2002] No.160 Technical Methods and Validation Guidelines for Removing Inactivated Virus from Blood Products[S].2002:1
[20] 王成彬.核酸检测用于确诊新型冠状病毒肺炎阳性率低的原因分析[J/OL].中华医学杂志,2020,100(00):E010-E010[2020-03-12].doi:10.3760/cma.j.cn112137-20200213-00280 WANG CB.Analysis of novel coronavirus pneumonia positive rate with nucleic acid detection[J/OL].Natl Med J China,2020,100(00):E010-E010[2020-03-12].doi:10.3760/cma.j.cn112137-20200213-00280
[21] 莫茜,秦炜,傅启华,等.正确认识新冠病毒核酸检测的影响因素[J/OL].中华检验医学杂志,2020,43[2020-03-12].http://rs.yiigle.com/yufabiao/1180120.htm.doi:10.3760/cma.j.issn.1009-8158.2020.0002.[Epub ahead of print] MO Q,QIN W,FU QH,et al.Correct understanding of the influencing factors of nucleic acid detection of new coronavirus[J/OL].Chin J Lab Med,2020,43[2020-03-12].http://rs.yiigle.com/yufabiao/1180120.htm.doi:10.3760/cma.j.issn.1009-8158.2020.0002.[Epub ahead of print]
[22] 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 MA J,HE P F,ZHAO C Y,et al.Development of SARS and MERS neutralization methods based on pseudoviruses[J].Chin J Virol,2019,35(2):189

欢迎阅读《药物分析杂志》!您是该文第 58位读者!

药物分析杂志 © 2009
地址:北京天坛西里2号 邮政编码:100050; 电子邮件:ywfx@nicpbp.org.cn

本系统由北京博思汇文数字科技有限公司设计开发 技术服务电话:400-921-9838