Comparative Analysis of RT-PCR and a Colloidal Gold Immunochromatographic Assay for SARS-CoV-2 Detection
Abstract
:1. Introduction
2. Materials and Methods
2.1. Sample Collection
2.2. Molecular RT-PCR Procedure
2.3. Colloidal Gold Immunochromatographic Assay (GICA)
2.4. Statistical Analysis
3. Results
3.1. Diagnostic Performance of the GICA Test Compared to RT-PCR
3.2. The Correlation Between the RT-PCR and GICA Results
3.3. Evaluation of Overall GICA Method Performance Against RT-PCR
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Abbreviations
SARS-CoV-2 | severe acute respiratory syndrome coronavirus 2 |
COVID-19 | coronavirus disease of 2019 |
N | nucleocapsid protein |
E | envelop protein |
mRT-PCR | multiplex reverse transcription polymerase chain reaction |
GICA | colloidal gold immunochromatographic assay |
POC | point of care |
Ct | cycle threshold |
NPV | negative predictive value |
PPV | positive predictive value |
ROC | receiver operating characteristic |
AUC | area under the curve |
CI | confidence intervals |
References
- Dong, T.; Wang, M.; Liu, J.; Ma, P.; Pang, S.; Liu, W.; Liu, A. Diagnostics and analysis of SARS-CoV-2: Current status, recent advances, challenges and perspectives. Chem. Sci. 2023, 14, 6149–6206. [Google Scholar] [CrossRef] [PubMed] [PubMed Central]
- Liu, D.; Cheng, Y.; Zhou, H.; Wang, L.; Fiel, R.H.; Gruenstein, Y.; Luo, J.J.; Singh, V.; Konadu, E.; James, K.; et al. Early Introduction and Community Transmission of SARS-CoV-2 Omicron Variant, New York, New York, USA. Emerg. Infect. Dis. 2023, 29, 371–380. [Google Scholar] [CrossRef] [PubMed] [PubMed Central]
- Zavascki, A.P. Urgent need for evaluation of point-of-care tests as an RT-PCR-sparing strategy for the diagnosis of Covid-19 in symptomatic patients. Epidemiol Infect. 2021, 149, e35. [Google Scholar] [CrossRef]
- Peeling, R.W.; Heymann, D.L.; Teo, Y.Y.; Garcia, P.J. Diagnostics for COVID-19: Moving from pandemic response to control. Lancet 2022, 399, 757–768. [Google Scholar] [CrossRef]
- Nichols, J.D.; Bogich, T.L.; Howerton, E.; Bjørnstad, O.N.; Borchering, R.K.; Ferrari, M.; Haran, M.; Jewell, C.; Pepin, K.M.; Probert, W.J.M.; et al. Strategic testing approaches for targeted disease monitoring can be used to inform pandemic decision-making. PLoS Biol. 2021, 19, e3001307. [Google Scholar] [CrossRef]
- Khan, M.H.; Becker, R.C. Bridging the gap: Strategies for recognizing and managing post-COVID conditions. Front. Med. 2024, 11, 1395420. [Google Scholar] [CrossRef]
- Hirabayashi, E.; Mercado, G.; Hull, B.; Soin, S.; Koshy-Chenthittayil, S.; Raman, S.; Huang, T.; Keerthisinghe, C.; Feliciano, S.; Dongo, A.; et al. Comparison of diagnostic accuracy of rapid antigen tests for COVID-19 compared to the viral genetic test in adults: A systematic review and meta-analysis. JBI Evid. Synth. 2024, 22, 1939–2002. [Google Scholar] [CrossRef]
- Wagenhäuser, I.; Knies, K.; Pscheidl, T.; Eisenmann, M.; Flemming, S.; Petri, N.; McDonogh, M.; Scherzad, A.; Zeller, D.; Gesierich, A.; et al. SARS-CoV-2 antigen rapid detection tests: Test performance during the COVID-19 pandemic and the impact of COVID-19 vaccination. EBioMedicine 2024, 109, 105394. [Google Scholar] [CrossRef]
- Drain, P.K.; Adams, A.K.; Kessler, L.; Thompson, M. A Call to Improve Usability, Accuracy, and Equity of Self-Testing for COVID-19 and Other Rapid Diagnostic Tests. Health Equity 2023, 7, 731–734. [Google Scholar] [CrossRef]
- Peña, M.; Ampuero, M.; Garcés, C.; Gaggero, A.; García, P.; Velasquez, M.S.; Luza, R.; Alvarez, P.; Paredes, F.; Acevedo, J.; et al. Performance of SARS-CoV-2 rapid antigen test compared with real-time RT-PCR in asymptomatic individuals. Int. J. Infect Dis. 2021, 107, 201–204. [Google Scholar] [CrossRef]
- Rankine-Wilson, L.; Oncken, T.; Basrewan, I.; Jeffery, C.; Pryce, T.M.; Wake, R.; Molan, A.A.; Paton, T.F.; Inglis, T.J. Lessons learned: Drive-through COVID-19 clinic testing during an adaptive epidemic response and a point-of-care test assessment of a computer-read rapid lateral flow immunoassay with fluorescence-based detection. J. Med. Microbiol. 2024, 73, 001875. [Google Scholar] [CrossRef] [PubMed]
- Chao, T.L.; Lee, W.H.; Hu, H.C.; Lin, Y.C.; Ho, S.Y.; Chen, H.H.; Chen, C.P.; Tsai, Y.M.; Fang, J.T.; Leong, S.M.; et al. Clinical assessment of SARS-CoV-2 infectivity by rapid antigen test compared with virus isolation. J. Clin. Virol. Plus. 2023, 3, 100133. [Google Scholar] [CrossRef] [PubMed]
- Ishige, T. Molecular biology of SARS-CoV-2 and techniques of diagnosis and surveillance. Adv. Clin. Chem. 2024, 118, 35–85. [Google Scholar] [CrossRef]
- Chiereghin, A.; Zagari, R.M.; Galli, S.; Moroni, A.; Gabrielli, L.; Venturoli, S.; Bon, I.; Rossini, G.; Saracino, I.M.; Pavoni, M.; et al. Orsola Polyclinic of Bologna COVID-19 Research Team. Recent Advances in the Evaluation of Serological Assays for the Diagnosis of SARS-CoV-2 Infection and COVID-19. Front. Public Health 2021, 8, 620222. [Google Scholar] [CrossRef] [PubMed] [PubMed Central]
- Jian, M.J.; Perng, C.L.; Chung, H.Y.; Chang, C.K.; Lin, J.C.; Yeh, K.M.; Chen, C.W.; Hsieh, S.S.; Pan, P.C.; Chang, H.T.; et al. Clinical assessment of SARS-CoV-2 antigen rapid detection compared with RT-PCR assay for emerging variants at a high-throughput community testing site in Taiwan. Int. J. Infect Dis. 2022, 115, 30–34. [Google Scholar] [CrossRef] [PubMed] [PubMed Central]
- Li, X.; Chen, H.; Wei, J.; Lv, N.; You, L. The evaluation of colloidal gold immunochromatographic assay (GICA) for rapid diagnosis of influenza A disease. Clin. Chem. Lab. Med. 2011, 49, 1533–1537. [Google Scholar] [CrossRef]
- Kunkel, D.; Stuenkel, M.; Sivaraj, L.B.; Colenda, C.C.; Pekarek, L.; Rennert, L. Predictive value of clinical symptoms for COVID-19 diagnosis in young adults. J. Am. Coll. Health. 2024, 72, 1006–1009. [Google Scholar] [CrossRef]
- Brajkovic, M.; Vukcevic, M.; Nikolic, S.; Dukic, M.; Brankovic, M.; Sekulic, A.; Popadic, V.; Stjepanovic, M.; Radojevic, A.; Markovic-Denic, L.; et al. The Predictive Value of Risk Factors and Prognostic Scores in Hospitalized COVID-19 Patients. Diagnostics 2023, 13, 2653. [Google Scholar] [CrossRef]
- Ishiguro, C.; Mimura, W.; Terada, J.; Matsunaga, N.; Ishiwari, H.; Hoshimoto, H.; Miyo, K.; Ohmagari, N. Development and Validation of Claims-based Algorithms for Identifying Hospitalized Patients With COVID-19 and Their Severity in 2020 and 2021. J. Epidemiol. 2024, 34, 485–492. [Google Scholar] [CrossRef]
- Ahuja, V.; Bowe, T.; Warnock, G.; Pitman, C.; Dwyer, D.E. Trends in SARS-CoV-2 cycle threshold (Ct) values from nucleic acid testing predict the trajectory of COVID-19 waves. Pathology 2024, 56, 710–716. [Google Scholar] [CrossRef]
- Liu, D.; Rodriguez, G.D.; Zhou, H.Y.; Cheng, Y.X.; Li, X.; Tang, W.; Prasad, N.; Chen, C.C.; Singh, V.; Konadu, E.; et al. SARS-CoV-2 Continuous Genetic Divergence and Changes in Multiplex RT-PCR Detection Pattern on Positive Retesting Median 150 Days after Initial Infection. Int. J. Mol. Sci. 2022, 23, 6254. [Google Scholar] [CrossRef] [PubMed] [PubMed Central]
- Green, D.A.; Zucker, J.; Westblade, L.F.; Whittier, S.; Rennert, H.; Velu, P.; Craney, A.; Cushing, M.; Liu, D.; Sobieszczyk, M.E.; et al. Clinical Performance of SARS-CoV-2 Molecular Tests. J. Clin. Microbiol. 2020, 58, e00995-20. [Google Scholar] [CrossRef] [PubMed] [PubMed Central]
- Phillips, M.C.; Quintero, D.; Wald-Dickler, N.; Holtom, P.; Butler-Wu, S.M. SARS-CoV-2 cycle threshold (Ct) values predict future COVID-19 cases. J. Clin. Virol. 2022, 150–151, 105153. [Google Scholar] [CrossRef] [PubMed] [PubMed Central]
- Westblade, L.F.; Brar, G.; Pinheiro, L.C.; Paidoussis, D.; Rajan, M.; Martin, P.; Goyal, P.; Sepulveda, J.L.; Zhang, L.; George, G.; et al. SARS-CoV-2 Viral Load Predicts Mortality in Patients with and without Cancer Who Are Hospitalized with COVID-19. Cancer Cell. 2020, 38, 661–671.e2. [Google Scholar] [CrossRef] [PubMed] [PubMed Central]
- Zhang, Y.; Xiao, W.; Kong, H.; Cheng, J.; Yan, X.; Zhang, M.; Wang, Q.; Qu, H.; Zhao, Y. A Highly Sensitive Immunochromatographic Strip Test for Rapid and Quantitative Detection of Saikosaponin d. Molecules 2018, 23, 338. [Google Scholar] [CrossRef] [PubMed] [PubMed Central]
- Zhang, Y.; Cao, P.; Lu, F.; Cheng, J.; Qu, H. Development of a Lateral Flow Immunochromatographic Strip for Rapid and Quantitative Detection of Small Molecule Compounds. J. Vis. Exp. 2021, 177, e62754. [Google Scholar] [CrossRef] [PubMed]
- Xu, M.; Lu, F.; Lyu, C.; Wu, Q.; Zhang, J.; Tian, P.; Xue, L.; Xu, T.; Wang, D. Broad-range and effective detection of human noroviruses by colloidal gold immunochromatographic assay based on the shell domain of the major capsid protein. BMC Microbiol. 2021, 21, 22. [Google Scholar] [CrossRef] [PubMed] [PubMed Central]
- Li, X.; Yin, Y.; Pang, L.; Xu, S.; Lu, F.; Xu, D.; Shen, T. Colloidal gold immunochromatographic assay (GICA) is an effective screening method for identifying detectable anti-SARS-CoV-2 neutralizing antibodies. Int. J. Infect Dis. 2021, 108, 483–486. [Google Scholar] [CrossRef] [PubMed] [PubMed Central]
- Tatliparmak, A.C.; Yilmaz, S.; Ak, R. Importance of receiver operating characteristic curve and decision curve analysis methods in clinical studies. Am. J. Emerg Med. 2023, 70, 196–197. [Google Scholar] [CrossRef]
- Burke, H.B. Use the Receiver Operating Characteristic to Assess Model Accuracy. JAMA Cardiol. 2023, 8, 998. [Google Scholar] [CrossRef]
- Ekström, J.; Åkerrén Ögren, J.; Sjöblom, T. Exact Probability Distribution for the ROC Area under Curve. Cancers 2023, 15, 1788. [Google Scholar] [CrossRef] [PubMed]
- Azami, M.; Jamshidbeigi, A.; Ghaleh, H.E.G.; Ranjbar, R. Biochemical, Serological and Immunological Tests for Diagnosis of Coronavirus Disease 2019 (COVID-19) Patients: A Comprehensive Systematic Review and Meta-analysis. Infect. Disord. Drug Targets 2023, 23, 42–65. [Google Scholar] [CrossRef] [PubMed]
- Alamri, S.S.; Alsaieedi, A.; Khouqeer, Y.; Afeef, M.; Alharbi, S.; Algaissi, A.; Alghanmi, M.; Altorki, T.; Zawawi, A.; Alfaleh, M.A.; et al. The importance of combining serological testing with RT-PCR assays for efficient detection of COVID-19 and higher diagnostic accuracy. PeerJ 2023, 11, e15024. [Google Scholar] [CrossRef]
- Lima, T.M.; Leal, D.M.; Ferreira, Z.C.; Souza, F.D.; de Oliveira, D.B.; Rocha-Vieira, E.; Martins, H.R.; Pereira, A.C.; Ferreira, L.F. Development and Optimization of a Cost-Effective Electrochemical Immunosensor for Rapid COVID-19 Diagnosis. Biosensors 2025, 15, 67. [Google Scholar] [CrossRef]
- Hassoun, M.R.; Kudlapur, N.T.; Chen, G.M.; Green-Ross, J.; Patel, A. Should rapid antigen tests be first-line for COVID-19 testing? Results of a prospective urban cohort study. BMC Infect. Dis. 2023, 23, 243. [Google Scholar] [CrossRef]
- Katzenschlager, S.; Bruemmer, L.E.; Schmitz, S.; Tolle, H.; Manten, K.; Gaeddert, M.; Erdmann, C.; Lindner, A.; Tobian, F.; Grilli, M.; et al. Comparing SARS-CoV-2 antigen-detection rapid diagnostic tests for COVID-19 self-testing/self-sampling with molecular and professional-use tests: A systematic review and meta-analysis. Sci. Rep. 2023, 13, 21913. [Google Scholar] [CrossRef]
- Rahimi, F.; Talebi Bezmin Abadi, A. Challenges of managing the asymptomatic carriers of SARS-CoV-2. Travel. Med. Infect. Dis. 2020, 37, 101677. [Google Scholar] [CrossRef] [PubMed] [PubMed Central]
- Tan, C.; Xiao, Y.; Meng, X.; Huang, X.; Li, C.; Wu, A. Asymptomatic SARS-CoV-2 infections: What do we need to know? Infect. Control Hosp. Epidemiol. 2021, 42, 114–115. [Google Scholar] [CrossRef] [PubMed] [PubMed Central]
Sample Number | Test Result | Grade | |
---|---|---|---|
1 | C:2613.290 | T:2.800 | − |
2 | C:2635.149 | T:3.208 | − |
3 | C:2679.448 | T:430.283 | + |
4 | C:2508.893 | T:372.090 | + |
5 | C:2596.239 | T:1185.282 | ++ |
6 | C:2551.200 | T:1082.041 | ++ |
7 | C:2403.839 | T:2521.277 | +++ |
8 | C:2568.612 | T:2624.942 | +++ |
9 | C:2312.836 | T:3312.668 | ++++ |
10 | C:2382.953 | T:3105.827 | ++++ |
RT-PCR Positive | RT-PCR Negative | Total | |
---|---|---|---|
GICA positive | 84 | 0 | 84 |
GICA negative | 33 | 12 | 45 |
Total | 117 | 12 | 129 |
Parameters | Values | 95% CI |
---|---|---|
Sensitivity | 71.8% | 63.6% to 79.9% |
Specificity | 100% | 100 to 100% |
Positive Predictive Value (PPV) | 100% | 100% to 100% |
Negative Predictive Value (NPV) | 26.7% | 13.7% to 39.6% |
Accuracy | 74.4% | 66.9% to 81.9% |
RT-PCR Ct Value Range | GICA Reading | No. of Samples Positive for RT-PCR * | No. of Samples Positive for GICA | Positive Rate |
---|---|---|---|---|
11.7–16.9 | 5 | 13 | 13 | 100% |
17.2–20.9 | 4 | 24 | 24 | 100% |
21.1–24.9 | 3 | 28 | 28 | 100% |
25.0–28.1 | 2 | 12 | 12 | 100% |
28.3–31.9 | 1 | 16 | 7 | 56.30% |
32.1–44.6 | 0 | 14 | 0 | 0% |
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to people or property resulting from any ideas, methods, instructions or products referred to in the content. |
© 2025 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Li, H.; Liu, D.; Zhou, Q.; Rodriguez, G.D.; Pietz, H.; Singh, V.; Konadu, E.; James, K.K.; Lui, C.; Shao, M.; et al. Comparative Analysis of RT-PCR and a Colloidal Gold Immunochromatographic Assay for SARS-CoV-2 Detection. Diagnostics 2025, 15, 1362. https://doi.org/10.3390/diagnostics15111362
Li H, Liu D, Zhou Q, Rodriguez GD, Pietz H, Singh V, Konadu E, James KK, Lui C, Shao M, et al. Comparative Analysis of RT-PCR and a Colloidal Gold Immunochromatographic Assay for SARS-CoV-2 Detection. Diagnostics. 2025; 15(11):1362. https://doi.org/10.3390/diagnostics15111362
Chicago/Turabian StyleLi, Hui, Dakai Liu, Qiang Zhou, George D. Rodriguez, Harlan Pietz, Vishnu Singh, Eric Konadu, Keither K. James, Calvin Lui, Mingyu Shao, and et al. 2025. "Comparative Analysis of RT-PCR and a Colloidal Gold Immunochromatographic Assay for SARS-CoV-2 Detection" Diagnostics 15, no. 11: 1362. https://doi.org/10.3390/diagnostics15111362
APA StyleLi, H., Liu, D., Zhou, Q., Rodriguez, G. D., Pietz, H., Singh, V., Konadu, E., James, K. K., Lui, C., Shao, M., Chen, J., Schreiner, A., Urban, C., Truong, J., Prasad, N., & Rodgers, W. H. (2025). Comparative Analysis of RT-PCR and a Colloidal Gold Immunochromatographic Assay for SARS-CoV-2 Detection. Diagnostics, 15(11), 1362. https://doi.org/10.3390/diagnostics15111362