加热灭活及4 ℃储存对血浆新型冠状病毒总抗体稳定性影响的研究  

Study on the SARS-CoV-2 antibody stability after heating-inactivation and 4℃preservation

作　　者：李爱敏 于思洋 董晓宇 童山石 林宝月 彭译漫 陈晓杭 张丽婷 张明霞[2] 张政 魏兰兰 Li Aiming;Yu Siyang;Dong Xiaoyu;Tong Shanshi;Ling Baoyue;Peng Yiman;Chen Xiaohang;Zhang Liting;Zhang Mingxia;Zhang Zheng;Wei Lanlan(Center Lab of Longhua Branch,Shenzhen People's Hospital,Shenzhen 518020,China;Institute of Hepatology,Shenzhen Third People's Hospital,Shenzhen 518112,China;Microbiology Department,Harbin Medical University,Harbin 150081,China;School of Medicine,Southern University of Science and Technology,Shenzhen 518055,China;The Genetics Laboratory,Longgang District Maternity&Child Healthcare Hospital of Shenzhen,Shenzhen 518172,China)

机构地区：[1]深圳市人民医院龙华分院检验科,518020 [2]深圳市第三人民医院肝病研究所,518112 [3]哈尔滨医科大学微生物学教研室,150081 [4]南方科技大学医学院,深圳518055 [5]深圳市龙岗区妇幼保健院中心实验室,518172

出　　处：《国际免疫学杂志》2021年第5期493-499,共7页International Journal of Immunology

基　　金：深圳市龙岗区医疗卫生科技计划项目(LGKCYLWS2020101)。

摘　　要：目的探讨严重急性呼吸综合征冠状病毒2(severe acute respiratory syndrome coronavirus 2,SARS-CoV-2)感染患者的血液标本经不同灭活和储存处理后,SARS-CoV-2总抗体稳定性的变化,从而论证以56℃,60 min加热作为患者血液标本安全有效的灭活方法的可行性。方法选取2020年2月24日至2020年3月30日深圳市第三人民医院收治的47例2019新型冠状病毒肺炎(corona virus disease 2019,COVID-19)确诊患者,另外选取同期因其他疾病入院的已排除患有COVID-19可能的33例住院患者。抽取两组研究对象的肘静脉乙二胺四乙酸(ethylene diamine tetraacetic acid,EDTA)抗凝血液,应用化学发光微粒子免疫检测法检测56℃,30 min灭活前后和60 min灭活前后血浆SARS-CoV-2总抗体水平,并在指定时间点检测SARS-CoV-2总抗体,进一步比较不同灭活处理方式及不同储存时间对SARS-CoV-2总抗体的影响。结果56℃,30 min灭活前后和60 min灭活前后,血浆样本中SARS-CoV-2总抗体检测阳性结果一致率分别为96.15%和100%,灭活前后总抗体样本临界指数(cut-off index,COI)值的Pearson相关系数分别为0.9965(95%CI:0.9920~0.9984,P<0.05)和0.9981(95%CI:0.9962~0.9990,P<0.05)。血浆样本经先离心再灭活处理与先灭活再离心处理,SARS-CoV-2总抗体阳性结果一致率为100%,两组总抗体COI值的Pearson相关系数为0.9995(95%CI:0.9950~0.9999,P<0.05)。未灭活样本4℃储存30 d;56℃,30 min或60 min灭活样本4℃储存10 d,SARS-CoV-2总抗体COI值均未出现显著性变化。结论56℃,30 min或60 min灭活处理血浆样本及灭活后4℃储存10 d,未灭活4℃储存30 d均不会对SARS-CoV-2总抗体的检测结果造成影响。Objective To investigate the stability changes of total antibody to severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)in infected patients'blood samples,after different inactivation treatments and storage conditions.Analyze the feasibility of 56℃inactivation for 60 min as a safe and effective treatment method for blood samples of patients.Methods Total of 47 patients diagnosed with 2019 corona virus disease 2019(COVID-19)from February 24,2020 to March 30,2020 in Shenzhen Third People's Hospital were selected.In addition,33 hospitalized patients who had been admitted to hospital due to other diseases in the same period and had excluded the possibility of suffering from COVID-19 were selected.Ethylene diamine tetraacetic acid(EDTA)anticoagulant blood of elbow vein was extracted from the two group of patients.The total antibody levels of SARS-CoV-2 were determined by chemiluminescent particle immunoassay before and after inactivation at 56℃,30 min or 60 min.The expression level of SARS-CoV-2 total antibody was detected at the specified time point,and the effects of different inactivation treatment methods and storage time on the total antibody of SARS-CoV-2 were further compared.Results Before and after inactivation at 56℃,30 min or 60 min,the coincident rate of positive results to SARS-CoV-2 total antibody in plasma samples was 96.15%and 100%respectively.Pearson correlation coefficients of total antibody cut-off index(COI)value before and after inactivation were 0.9965(95%CI:0.9920~0.9984,P<0.05)and 0.9981(95%CI:0.9962~0.9990,P<0.05)respectively.The coincident rate of positive results to SARS-CoV-2 total antibody was 100%,whether the plasma sample is centrifuged first and then inactivated or inactivated first and then centrifuged.The Pearson correlation coefficient of total antibody COI value between the two groups was 0.9995(95%CI:0.9950~0.9999,P<0.05).The total antibody COI value of SARS-CoV-2 did not change significantly when the uninactivated samples were stored at 4℃for 30 d,or the samples wer

关 键 词：新型冠状病毒 抗体稳定性 病毒灭活 离心 储存时间 

分 类 号：R392-33[医药卫生—免疫学]