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[Effect of heat inactivation of blood samples on the efficacy of three detection methods of SARS-CoV-2 antibodies].

ABSTRACT: OBJECTIVE:To evaluate the effects of heat inactivation of blood samples at 56? for 30 min on the results of SARS-CoV-2 antibody detection using different methods. METHODS:This retrospective study was conducted in 11 patients with established diagnosis of COVID-19 and 10 patients with diseases other than COVID- 19 in our hospital. We collected samples of serum, plasma and whole blood from each patient between February, 12 and 18, 2020, and with a double- blind design, the samples were examined for SARS-CoV-2 antibodies before and after heat inactivation at 56 ? for 30 min. In all the samples, the total SARS-CoV-2 antibodies were detected using immunochromatography, and the IgM antibodies were detected using fluorescence immunochromatography; the IgM and IgG antibodies in the serum and plasma samples detected with chemiluminescence immunoassay. We compared the detection results and analyzed the correlation of semi-quantitative detection results of IgM and IgG antibodies before and after heat inactivation of the samples. RESULTS:With immuno-chromatography, the coincidence rate of the total SARS-CoV-2 antibody detection before and after heat inactivation of the serum and plasma samples was 90.0% in COVID-19 cases and 100.0% in the negative cases, resulting in a total coincidence rate 95.2%; for the whole blood samples, the total coincidence rates of the total SARS-CoV-2 antibodies were 100.0%. For detection of IgM antibodies in the serum, plasma and whole blood samples using fluorescence immunochromatography, the coincidence rates in SARS-CoV-2-positive and negative cases and the total coincidence rate before and after inactivation were 100.0%, 0 and 47.6%, respectively. For detection of serum IgM and IgG antibodies and plasma IgG antibodies with chemiluminescence immunoassay, the coincidence rates in SARS-CoV-2-positive and negative cases and the total coincidence rate before and after inactivation were all 100.0%, and the total coincidence rate of plasma IgM antibodies was 95.2%. Pearson correlation analysis of the semi-quantitative results of IgM and IgG detection in the serum and plasma samples showed a correlation coefficient of 0.9999 (95%CI: 0.9998-1.000, P < 0.001) between the results before and after sample inactivation. CONCLUSIONS:Heat inactivation of blood samples at 56 ? for 30 min does not obviously affect the results of immunochromatography and chemiluminescent immunoassay for detection of SARS-COV-2 antibodies but can reduce the risk of infection for the operators. Heat-inactivated samples can not be used in fluorescence immunochromatography for SARS-CoV-2 antibody detection.

SUBMITTER: Xue X

PROVIDER: S-EPMC7167315 | biostudies-literature | 2020 Mar

REPOSITORIES: biostudies-literature

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[Effect of heat inactivation of blood samples on the efficacy of three detection methods of SARS-CoV-2 antibodies].

Xue Xiongyan X Zhu Changlin C Huang Shaozhen S Pan Lianhua L Xu Jianhua J Li Weixuan W

Nan fang yi ke da xue xue bao = Journal of Southern Medical University 20200301 3

<h4>Objective</h4>To evaluate the effects of heat inactivation of blood samples at 56℃ for 30 min on the results of SARS-CoV-2 antibody detection using different methods.<h4>Methods</h4>This retrospective study was conducted in 11 patients with established diagnosis of COVID-19 and 10 patients with diseases other than COVID- 19 in our hospital. We collected samples of serum, plasma and whole blood from each patient between February, 12 and 18, 2020, and with a double- blind design, the samples w ...[more]

PMID: 32376571

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Project description:Assessing the levels of serum IgG antibodies is widely used to measure immunity to influenza and the new severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) after natural infection or vaccination with specific vaccines, as well as to study immune responses to these viruses in animal models. For safety reasons, sometimes serum specimens collected from infected individuals are subjected to heat inactivation at 56 °C to reduce the risk of infecting personnel during serological studies. However, this procedure may affect the level of virus-specific antibodies, making the results of antibody immunoassays uninterpretable. Here, we evaluated the effect of the heat inactivation of human, ferret and hamster serum samples on the binding of IgG antibodies to the influenza and SARS-CoV-2 antigens. For this, serum samples of naive and immune hosts were analyzed in three variants: (i) untreated sera, (ii) heated at 56 °C for 1 h, and (iii) treated with receptor-destroying enzyme (RDE). The samples were studied through an in-house enzyme-linked immunosorbent assay (ELISA) using whole influenza virus or recombinant proteins corresponding to nucleocapsid (N) protein and the receptor-binding domain of SARS-CoV-2 Spike (RBD) as antigens. We demonstrated that the heat inactivation of the naive serum samples of various hosts can lead to false-positive results, while RDE treatment abolished the effect of the non-specific binding of IgG antibodies to the viral antigens. Furthermore, RDE also significantly decreased the level of virus-specific IgG antibodies in SARS-CoV-2 and influenza-immune sera of humans and animals, although it is unknown whether it actually removes true virus-specific IgG antibodies or only non-specifically binding artifacts. Nevertheless, we suggest that the RDE treatment of human and animal sera may be useful in preventing false-positive results in various immunoassays, while also neutralizing infectious virus, since the standard protocol for the use of RDE also includes heating the sample at 56 °C.

Dataset Information

[Effect of heat inactivation of blood samples on the efficacy of three detection methods of SARS-CoV-2 antibodies].

Publications

[Effect of heat inactivation of blood samples on the efficacy of three detection methods of SARS-CoV-2 antibodies].

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