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Abstract

Several studies have investigated the effect of repeated freeze–thaw (F/T) cycles on RNA detection for severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). However, no data are available regarding the effect of repeated F/T cycles on SARS-CoV-2 antibody detection in serum. We investigated the effect of multiple F/T cycles on anti-SARS-CoV-2 IgG detection using an ELISA test targeting the nucleocapsid antibodies. Ten positive and 1 negative SARS-CoV-2 IgG sera from 11 participants, in replicates of 5, were subjected to a total of 16 F/T cycles and stored at 4 °C until tested by ELISA. Statistical analysis was performed to test for F/T cycle effect. None of the 10 positive sera became negative after 16 F/T cycles. There was no significant difference in the OD average reading between the first and last F/T cycles, except for one serum with a minimal decline in the OD. The random effect linear regression of log (OD) on the number of cycles showed no significant trend, with a slope consistent with zero (B=−0.0001; 95 % CI −0.0008; 0.0006; -value=0.781). These results suggest that multiple F/T cycles had no effect on the ability of the ELISA assay to detect SARS-CoV-2 IgG antibodies.

Keyword(s): antibodies , COVID-19 , detection , freeze , IgG , SARS-CoV-2 and thaw
Funding
This study was supported by the:
  • Qatar University (Award M-QJRC-2020-5 grant.)
    • Principle Award Recipient: LaithJ. Abu Raddad
  • Qatar University (Award QUERG-CMED-2020-2)
    • Principle Award Recipient: GheyathNasrallah
  • Qatar National Research Fund (Award RRC-2-032)
    • Principle Award Recipient: GheyathNasrallah
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License. The Microbiology Society waived the open access fees for this article.
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/content/journal/jmm/10.1099/jmm.0.001402
2021-08-06
2024-12-05
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References

  1. Castejon MJ, Yamashiro R, Oliveira CC, Oliveira EL, Silveira EP et al. Effect of multiple freeze-thaw cycles on the stability of positive anti-treponemal serum samples. Jornal Brasileiro de Patologia e Medicina Laboratorial 2017; 53:246–251 [View Article]
    [Google Scholar]
  2. Castro AR, Jost HA. Effect of multiple freeze and thaw cycles on the sensitivity of IGG and IGM immunoglobulins in the sera of patients with syphilis. Sex Transm Dis 2013; 40:870–871 [View Article]
    [Google Scholar]
  3. Guo G-H, Dong J, Yuan X-H, Dong Z-N, Tian Y-P. Clinical evaluation of the levels of 12 cytokines in serum/plasma under various storage conditions using evidence biochip arrays. Mol Med Rep 2013; 7:775–780 [View Article]
    [Google Scholar]
  4. Cuhadar S, Koseoglu M, Atay A, Dirican AJB. The effect of storage time and freeze-thaw cycles on the stability of serum samples. Biochem Med (Zagreb) 2013; 23:70–77 [View Article]
    [Google Scholar]
  5. Miller MA, Rodrigues MA, Glass MA, Singh SK, Johnston KP et al. Frozen-state storage stability of a monoclonal antibody: aggregation is impacted by freezing rate and solute distribution. J Pharm Sci 2013; 102:1194–1208 [View Article]
    [Google Scholar]
  6. Paltiel L, Rønningen KS, Meltzer HM, Baker SV, Hoppin Jajc. Evaluation of Freeze Thaw Cycles on stored plasma in the Biobank of the Norwegian Mother and Child Cohort study. Cell Preserv Technol 2008; 6:223–230 [View Article]
    [Google Scholar]
  7. Maelegheer K, Devreese KMJ. The impact of repeated freeze-thaw cycles on antiphospholipid antibody titer. Res Pract Thromb Haemost 2018; 2:366–369 [View Article]
    [Google Scholar]
  8. Pinsky NA, Huddleston JM, Jacobson RM, Wollan PC, Poland GA et al. Effect of multiple freeze-thaw cycles on detection of measles, mumps, and rubella virus antibodies. Clin Diagn Lab Immunol 2003; 10:19–21 [View Article]
    [Google Scholar]
  9. Petrakis NL. Biologic banking in cohort studies, with special reference to blood. Natl Cancer Inst Monogr 1985; 67:193–198 [PubMed]
    [Google Scholar]
  10. Younes S, Younes N, Shurrab F, Nasrallah GK. Severe acute respiratory syndrome coronavirus-2 natural animal reservoirs and experimental models: systematic review. Rev Med Virol 2020e2196 [View Article]
    [Google Scholar]
  11. Li L, Li X, Guo Z, Wang Z, Zhang K et al. Influence of storage conditions on SARS-COV-2 nucleic acid detection in throat swabs. J Infect Dis 2020; 222:203–205 [View Article]
    [Google Scholar]
  12. Stohr JJJM, Wennekes M, van der Ent M, Diederen BMW, Kluytmans-van den Bergh MFQ et al. Clinical performance and sample freeze-thaw stability of the cobas®6800 SARS-CoV-2 assay for the detection of SARS-CoV-2 in oro-/nasopharyngeal swabs and lower respiratory specimens. J Clin Virol 2020; 133:104686 [View Article]
    [Google Scholar]
  13. Al-Thani MH, Farag E, Bertollini R, Al Romaihi HE, Abdeen S et al. Seroprevalence of SARS-CoV-2 infection in the craft and manual worker population of Qatar. medRxiv 2020
    [Google Scholar]
  14. Yassine HM, Al-Jighefee H, Al-Sadeq DW, Dargham SR, Younes SN et al. Performance evaluation of five ELISA kits for detecting anti-SARS-COV-2 IgG antibodies. Int J Infect Dis 2021; 102:181–187 [View Article]
    [Google Scholar]
  15. Demir M, Cevahir N. Does multiple freezing and thawing cycles of serum affect the detection of anti-nuclear antibodies and anti-neutrophil cytoplasmic antibodies by indirect immunofluorescent method?. Biomed Res 2014; 25:
    [Google Scholar]
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