Follow-up testing of borderline SARS-CoV-2 patients by rRT-PCR allows early diagnosis of COVID-19

Joost Boeckmans, Reinoud Cartuyvels, Petra Hilkens, Liesbeth Bruckers, Koen Magerman, Luc Waumans, Marijke Raymaekers, Joost Boeckmans, Reinoud Cartuyvels, Petra Hilkens, Liesbeth Bruckers, Koen Magerman, Luc Waumans, Marijke Raymaekers

Abstract

Detection of SARS-CoV-2 RNA in nasopharyngeal samples using the real-time reverse transcription polymerase chain reaction (rRT-PCR) is the gold standard for diagnosing COVID-19. Determination of SARS-CoV-2 RNA by rRT-PCR sometimes results in an inconclusive test result due to a high cycle threshold-value. We retrospectively analyzed 30,851 SARS-CoV-2 rRT-PCR test results. Borderline positivity was considered as the presence of ≤25 viral copies per milliliter, while no amplification was considered as a negative test result. Of all test results, 204 were answered as borderline, of which 107 were accompanied by a follow-up test within 96 hours. Of the 107 follow-up samples, 10 (9.35%) were found positive for SARS-CoV-2. COVID-19 symptoms were not predictive for testing positive in the follow-up test. The positive SARS-CoV-2 samples in the follow-up group represented 0.92% of all positive test results, highlighting the need for retesting and increased hygienic measures for borderline SARS-CoV-2 patients [NCT04636294].

Keywords: Borderline; COVID-19; Molecular diagnostics; SARS-CoV-2; rRT-PCR.

Copyright © 2021 Elsevier Inc. All rights reserved.

Figures

Fig. 1
Fig. 1
Description of the patient cohort. (A) Organigram of positive, negative and borderline SARS-CoV-2 rRT-PCR test results. (B) From a total of 204 SARS-CoV-2 borderline samples, 18 were preceded by a positive test result.
Fig. 2
Fig. 2
Follow-up of borderline SARS-CoV-2 rRT-PCR results. (A) Ten out of 107 patients (9.35%) tested positive within 96 hours after a borderline SARS-CoV-2 test result. (B) Follow-up viral loads were significantly higher in SARS-CoV-2-positive patients (Wilcoxon signed rank test). (C) Patients with a positive SARS-CoV-2 test result within 96 hours after a borderline test result, did not experience more often COVID-19 symptoms upon first presentation compared to patients that turned out SARS-CoV-2 negative in the follow-up test (Fischer's exact test).

References

    1. Boeckmans J, Rodrigues RM, Demuyser T, Piérard D, Vanhaecke T, Rogiers V. COVID‑19 and drug‑induced liver injury : a problem of plenty or a petty point ? Arch Toxicol. 2020;94:1367–1369. doi: 10.1007/s00204-020-02734-1.
    1. Botta M, Tsonas AM, Pillay J, Boers LS, Algera AG, Bos LDJ, et al. Ventilation management and clinical outcomes in invasively ventilated patients with COVID-19 (PRoVENT-COVID): a national, multicentre, observational cohort study. Lancet Respir Med. 2020;19:1–10. doi: 10.1016/s2213-2600(20)30459-8.
    1. Cento V, Colagrossi L, Nava A, Lamberti A, Senatore S, Travi G, et al. Persistent positivity and fluctuations of SARS-CoV-2 RNA in clinically-recovered COVID-19 patients. J Infect. 2020;81:90–92. doi: 10.1016/j.jinf.2020.06.024.
    1. Cevik M, Tate M, Lloyd O, Maraolo AE, Schafers J, Ho A. SARS-CoV-2, SARS-CoV-1 and MERS-CoV viral load dynamics, duration of viral shedding and infectiousness: a living systematic review and meta-analysis. Lancet Microbe. 2020;5247:1–10. doi: 10.2139/ssrn.3677918.
    1. Cheng HY, Jian SW, Liu DP, Ng TC, Huang WT, Lin HH. Contact tracing assessment of COVID-19 transmission dynamics in taiwan and risk at different exposure periods before and after symptom onset. JAMA Intern Med. 2020;180:1156–1163. doi: 10.1001/jamainternmed.2020.2020.
    1. Chu DK, Akl EA, Duda S, Solo K, Yaacoub S, Schünemann HJ, et al. Physical distancing, face masks, and eye protection to prevent person-to-person transmission of SARS-CoV-2 and COVID-19: a systematic review and meta-analysis. Lancet. 2020;395:1973–1987. doi: 10.1016/S0140-6736(20)31142-9.
    1. D'Ardes D, Boccatonda A, Rossi I, Pontolillo M, Cocco G, Schiavone C, et al. Long-term positivity to SARS-CoV-2: a clinical case of COVID-19 with persistent evidence of infection. Eur J Case Rep Intern Med. 2020;7 doi: 10.12890/2020_001707.
    1. Dabouh E, Lázaro-Perona F, Romero-gómez M, Mingorance J, Garciá-Rodriguez J. Ct values from SARS-CoV-2 diagnostic PCR assays should not be used as direct estimates of viral load. J Infect. 2020;21 doi: 10.1016/j.jinf.2020.10.017. In press.
    1. Dan JM, Mateus J, Kato Y, Hastie KM, Faliti CE, Ramirez SI, et al. Immunological memory to SARS-CoV-2 assessed for up to 8 months after infection. Science. 2021;4063:1–23. doi: 10.1101/2020.11.15.383323.
    1. Drew RJ, Donnell SO, Leblanc D, Mcmahon M, Natin D. The importance of cycle threshold values in interpreting molecular tests for SARS-CoV-2. Diagn Microbiol Infect Dis. 2020;98:1–3. doi: 10.1016/j.diagmicrobio.2020.115130.
    1. Fiorentini S, Messali S, Zani A, Caccuri F, Giovanetti M, Ciccozzi M, et al. First detection of SARS-CoV-2 spike protein N501 mutation in Italy in August, 2020. Lancet Infect Dis. 2021;3099:3099. doi: 10.1016/s1473-3099(21)00007-4.
    1. Fischer M, Renevey N, Thür B, Hoffmann D, Beer M, Hoffmann B. Efficacy assessment of nucleic acid decontamination reagents used in molecular diagnostic laboratories. PLoS One. 2016;11:1–9. doi: 10.1371/journal.pone.0159274.
    1. Grant MC, Geoghegan L, Arbyn M, Mohammed Z, McGuinness L, Clarke EL, et al. The prevalence of symptoms in 24,410 adults infected by the novel coronavirus (SARS-CoV-2; COVID-19): a systematic review and meta-analysis of 148 studies from 9 countries. PLoS One. 2020;15 doi: 10.1371/journal.pone.0234765.
    1. He W, Yi GY, Zhu Y. Estimation of the basic reproduction number, average incubation time, asymptomatic infection rate, and case fatality rate for COVID-19: meta-analysis and sensitivity analysis. J Med Virol. 2020;92:2543–2550. doi: 10.1002/jmv.26041.
    1. Jamal AJ, Mozafarihashjin M, Coomes E, Powis J, Li AX, Paterson A, et al. Sensitivity of nasopharyngeal swabs and saliva for the detection of severe acute respiratory syndrome Coronavirus 2. Clin Infect Dis. 2020;27:9–11. doi: 10.1093/cid/ciaa848.
    1. Khalili M, Karamouzian M, Nasiri N, Javadi S, Mirzazadeh A, Sharifi H. Epidemiological characteristics of COVID-19: a systematic review and meta-analysis. Epidemiol Infect. 2020;148:1–17. doi: 10.1017/S0950268820001430.
    1. Lan L, Xu D, Ye G, Xia C, Wang S, Li Y, et al. Positive RT-PCR test results in patients recovered from COVID-19. J Am Med Assoc. 2020;323:1502–1503. doi: 10.1001/jama.2020.2783.
    1. Li N, Wang X, Lv T. Prolonged SARS-CoV-2 RNA shedding: not a rare phenomenon. J Med Virol. 2020;92:2286–2287. doi: 10.1002/jmv.25952.
    1. Lohse S, Pfuhl T, Berkó-Göttel B, Rissland J, Geißler T, Gärtner B, et al. Pooling of samples for testing for SARS-CoV-2 in asymptomatic people. Lancet Infect Dis. 2020;20:1231–1232. doi: 10.1016/S1473-3099(20)30362-5.
    1. Lotfi M, Hamblin MR, Rezaei N. COVID-19: Transmission, prevention, and potential therapeutic opportunities. Clin Chim Acta. 2020;508:254–266. doi: 10.1016/j.cca.2020.05.044.
    1. Rao SN, Manissero D, Steele VR, Pareja J. A narrative systematic review of the clinical utility of cycle threshold values in the context of COVID-19. Infect Dis Ther. 2020;9:573–586. doi: 10.1007/s40121-020-00324-3.
    1. Raymaekers M, Smets R, Maes B, Cartuyvels R. Checklist for optimization and validation of real-time PCR assays. J Clin Lab Anal. 2009;23:145–151. doi: 10.1002/jcla.20307.
    1. Romano M, Ruggiero A, Squeglia F, Maga G, Berisio R. A structural view of SARS-CoV-2 RNA replication machinery: rna synthesis, proofreading and final capping. Cells. 2020;9:1267. doi: 10.3390/cells9051267.
    1. Ruiz-Villalba A, van Pelt-Verkuil E, Gunst QD, Ruijter JM, van den Hoff MJ. Amplification of nonspecific products in quantitative polymerase chain reactions (qPCR) Biomol Detect Quantif. 2017;14:7–18. doi: 10.1016/j.bdq.2017.10.001.
    1. Tahamtan A, Ardebili A. Real-time RT-PCR in COVID-19 detection: issues affecting the results. Expert Rev Mol Diagn. 2020;20:453–454. doi: 10.1080/14737159.2020.1757437.
    1. Walsh KA, Jordan K, Clyne B, Rohde D, Drummond L, Byrne P, et al. SARS-CoV-2 detection, viral load and infectivity over the course of an infection. J Infect. 2020;81:357–371. doi: 10.1016/j.jinf.2020.06.067.
    1. Wang J, Hang X, Wei B, Li D, Chen F, Liu W, et al. Persistent SARS-COV-2 RNA positivity in a patient for 92 days after disease onset: A case report. Medicine (Baltimore) 2020;99:e21865. doi: 10.1097/MD.0000000000021865.
    1. Zhang ZL, Hou YL, Li DT, Li FZ. Diagnostic efficacy of anti-SARS-CoV-2 IgG/IgM test for COVID-19: a meta-analysis. J Med Virol. 2020 doi: 10.1002/jmv.26211. In press.

Source: PubMed

スポンサーと協力者

医学的状態

薬物療法

3
購読する