Detection of SARS-CoV-2 by antigen ELISA test is highly swayed by viral load and sample storage condition,Expert Review of Anti-infective Therapy

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Detection of SARS-CoV-2 by antigen ELISA test is highly swayed by viral load and sample storage condition
Expert Review of Anti-infective Therapy ( IF 4.2 ) Pub Date : 2021-09-11 , DOI: 10.1080/14787210.2021.1976144
Nihad Adnan ₁ , Shahad Saif Khandker ₂ , Ahsanul Haq ₂ , Mousumi Akter Chaity ₁ , Abdul Khalek ₃ , Anawarul Quader Nazim ₃ , Taku Kaitsuka ₄ , Kazuhito Tomizawa ₅ , Masayasu Mie ₆ , Eiry Kobatake ₆ , Sohel Ahmed ₇ , Nor Azlina A Ali ₈ , Mohib Ullah Khondoker ₉ , Mainul Haque ₁₀ , Mohd Raeed Jamiruddin ₁₁

Affiliation

Department of Microbiology, Jahangirnagar University, Dhaka, Bangladesh.
Department of Research and Development, Gonoshasthaya-RNA Molecular Diagnostic & Research Center, Dhaka, Bangladesh.
Department of Diagnostic Laboratory, Enam Medical College and Hospital, Dhaka, Bangladesh.
Department of Pharmaceutical Sciences, School of Pharmacy, International University of Health and Welfare, Fukuoka, Japan.
Department of Molecular Physiology, Faculty of Life Sciences, Kumamoto University, Kumamoto, Japan.
Department of Life Science and Technology,School of Life Science and Technology, Tokyo Institute of Technology, Yokohama, Japan.
Department of Biochemistry and Molecular Biology, Jahangirnagar University, Dhaka, Bangladesh.
Department of Physical Rehabilitation Sciences, Kulliyyah of Allied Health Sciences, International Islamic University Malaysia, Kuantan, Malaysia.
Department of Community Medicine, Gonoshasthaya Samaj Vittik Medical College, Dhaka, Bangladesh.
The Unit of Pharmacology, Faculty of Medicine and Defence Health Universiti Pertahanan, Nasional Malaysia (National Defence University of Malaysia), Kuala Lumpur, Malaysia.
Department of Pharmacy, Brac University, Dhaka, Bangladesh.

ABSTRACT

Background

Rapid increase in COVID-19 suspected cases has rendered disease diagnosis challenging, mainly depending upon RT-qPCR. Reliable, rapid, and cost-effective diagnostic assays that complement RT-qPCR should be introduced after thoroughly evaluating their performance upon various disease phases, viral load, and sample storage conditions.

Objective

We investigated the correlation of cycle threshold (Ct) value, which implies the viral load and infection phase, and the storage condition of the clinical specimen with the diagnosis of SARS-CoV-2 through our newly developed in-house rapid enzyme-linked immunosorbent assay (ELISA) system.

Method

Naso-oropharyngeal samples of 339 COVID-19 suspected cases were collected and evaluated through RT-qPCR that were stored up to 30 days in different conditions (i.e. −80°C, −20°C and initially at 4°C followed by −80°C). The clinical specimens were evaluated with our in-house ELISA system after finalizing the assay method through checkerboard assay and minimizing the signal/noise ratio.

Result

The ELISA system showed the highest sensitivity (92.9%) for samples with Ct ≤30 and preserving at −80°C temperature. The sensitivity reduced proportionally with increasing Ct value and preserving temperature. However, the specificity ranged between 98.3% and 100%.

Conclusion

The results indicate the necessity of early infection phase diagnosis and lower temperature preservation of samples to perform rapid antigen ELISA tests.

中文翻译：

通过抗原 ELISA 检测对 SARS-CoV-2 的检测受病毒载量和样本储存条件的影响很大

抽象的

背景

COVID-19 疑似病例的快速增加给疾病诊断带来了挑战，主要依靠 RT-qPCR。在彻底评估 RT-qPCR 在不同疾病阶段、病毒载量和样品储存条件下的性能后，应引入可靠、快速且经济有效的诊断检测方法来补充 RT-qPCR。

客观的

我们通过我们新开发的内部快速酶联免疫吸附剂，研究了循环阈值 (Ct) 值（暗示病毒载量和感染阶段）以及临床标本的储存条件与 SARS-CoV-2 诊断的相关性测定（ELISA）系统。

方法

收集了 339 例 COVID-19 疑似病例的鼻口咽样本，并通过 RT-qPCR 进行评估，这些样本在不同条件下保存长达 30 天（即 -80°C、-20°C，最初为 4°C，随后为 -80°C）。 ℃）。通过棋盘测定最终确定测定方法并最小化信噪比后，使用我们的内部 ELISA 系统对临床标本进行评估。