Comparison of the Plaque Assay and 50% Cell Culture Infectious Dose Assay Methods for Quantification Measuring SARS-CoV-2 Wuhan-Hu-1 Strain Infectivity

Main Article Content

Sakalin Trisiriwanich
Sompong Sapsutthipas
Kornnika Kullabutr
Ashma Yuso
Supaporn Phumiamorn

Abstract

Background: The Corona Virus Disease 2019 (COVID-19), which causes from the Severe Acute Respiratory Syndrome Corona Virus (SAR-CoV-2), is an emerging disease that several organizations have tried to develop vaccines against the viruses and therapeutic medicines, including specific anti-viral drugs. Therefore, viral cultivation and quantification are necessary for the laboratory testing. Most of common methods to quantify the viral titer are the Plaque and CCID50 assays.


Objective: To compare the quantity of the SAR-CoV-2 and the consumption resources test between the plaque and CCID50 assays (original strain).


Method: This study was an experimental research from March to June 2020 by comparing the quantification of the SAR-CoV-2 between both methods using the same virus set at 80 tests in the same period.


Result: The results showed that the geometric means of viral titers determined by the plaque and CCID50 assays were 5.97 log PFU/ml and 6.00 log CCID50/ml, respectively. The percentages of the coefficient of variance of the methods were 4.09 and 3.29, respectively. The statistical analysis by the Paired-t test showed no statistically significant difference between these two methods at a 95% confidence interval. In addition, the ratio of viral titers of the mean over the geometric mean ranged between 0.87 and 1.13. However, the CCID50 assay was faster by requiring only 4 days of testing and more cost-effective by using fewer materials, equipment, and medium.


Conclusion: The determination of SARS-CoV-2 virus counting by the plague counting method and the 50% cell culture infection test showed no difference in results, but testing for 50% infection in cell cultures was faster and more cost-effective. It revealed that both methods can be used to quantify the SARS-CoV-2 titer, which can be applied to detect immune responses in COVID-19 patients as well as in COVID-19 vaccine testing further in animals and humans.

Article Details

How to Cite
1.
Trisiriwanich S, Sapsutthipas S, Kullabutr K, Yuso A, Phumiamorn S. Comparison of the Plaque Assay and 50% Cell Culture Infectious Dose Assay Methods for Quantification Measuring SARS-CoV-2 Wuhan-Hu-1 Strain Infectivity. TFDJ [Internet]. 2023 Aug. 3 [cited 2024 Apr. 18];30(2):33-48. Available from: https://he01.tci-thaijo.org/index.php/fdajournal/article/view/265170
Section
Research Article
Author Biographies

Sakalin Trisiriwanich, Institute of Biological Products, Department of Medical Sciences

Institute of Biological Products, Department of Medical Sciences, Ministry of Public Health Tivanon Rd, Nonthaburi 11000, Thailand

Sompong Sapsutthipas, Institute of Biological Products, Department of Medical Sciences

Institute of Biological Products, Department of Medical Sciences, Ministry of Public Health Tivanon Rd, Nonthaburi 11000, Thailand

Kornnika Kullabutr, Institute of Biological Products, Department of Medical Sciences

Institute of Biological Products, Department of Medical Sciences, Ministry of Public Health Tivanon Rd, Nonthaburi 11000, Thailand

Ashma Yuso, Institute of Biological Products, Department of Medical Sciences

Institute of Biological Products, Department of Medical Sciences, Ministry of Public Health Tivanon Rd, Nonthaburi 11000, Thailand

Supaporn Phumiamorn, Institute of Biological Products, Department of Medical Sciences

Institute of Biological Products, Department of Medical Sciences

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