Overview of the SARS-CoV-2 (Coronavirus) Detection Process

In December 2019 the novel SARS-CoV-2 (formerly named 2019-nCoV or commonly named coronavirus) was identified in Wuhan, the capital of China’s province Hubei. Observations show that the virus is capable of spreading from person to person and causes COVID-19 – an acute respiratory disease with incubation periods estimated between 2-14 days. Based on its rapid spreading with more than 87,000 confirmed cases by the beginning of March 2020 the World Health Organization declared the outbreak a public health emergency of international concern.

Important facts:

• SARS-CoV-2: (+) ss-RNA virus
• Related to SARS-CoV and MERS-CoV
• Detection based on basis of PCR

Testing induced for:

• People with acute respiratory symptoms AND stay in risk area
• Symptomatic people that had contact to people of confirmed COVID-19
• Negative results despite of strong suspicions for infection require new sampling and PCR analysis

Complete Detection Workflow

Analytik Jena suggests the complete workflow from sampling until the final detection of the virus. This article is giving you detailed information for sampling, extraction and detection.

1. Sample Preparation

According to WHO¹:

• Respiratory material* (nasopharyngeal and oropharyngeal swab in ambulatory patients and sputum (if produced) and/or endotracheal aspirateor bronchoalveolar lavage in patients with more severe respiratory disease)
• Serum for serological testing, acute sample and convalescent sample (this is additional to respiratory materials and can support the identification of the true agent, once serologic assay is available)

*Modifiable with information on whether upper or lower respiratory material is better for coronavirus detection.
¹www.who.int/publications-detail/laboratory-testing-for-2019-novel-coronavirus-in-suspected-human-cases-20200117

2. Nucleic Acid Extraction

Analytik Jena offers a choice of extraction platforms and related extraction kits for extraction of viral RNA from swabs, cell free body fluids (e.g. serum, plasma, liquor) and more. The product line innuPREP AniPath allows the simultanous extraction of both bacterial and viral DNA and/or RNA from respective starting materials. A detailed overview is shown in the table below.

andling	Sample troughput	Extraction platform		Extraction kit
		Name	Key features
Manual	Low	–	Up to 12 samples / 25 min	innuPREP Virus DNA/RNA Kit innuPREP Virus RNA Kit
Automated	Middle	InnuPure C16 touch	Up to 16 samples / 31 – 84 min Extraction platform only	innuPREP AniPath DNA/RNA Kit – IPC16 innuPREP Virus DNA/RNA Kit – IPC16 (MDX)* innuPREP Virus DNA/RNA Kit – IPC16
	High	CyBio FeliX	Up to 96 samples / 62 – 73 min Extraction platform + liquid handling (e.g. preparation of PCR)	innuPREP AniPath DNA/RNA Kit – FX** innuPREP Virus DNA/RNA Kit – FX
	High	KingFisher Flex	Up to 96 samples / 60 min Extraction platform only	innuPREP AniPath DNA/RNA Kit – KFFLX innuPREP RNA Virus PLUS Kit (MDX) – KFFLX * innuPREP RNA Virus PLUS Kit – KFFLX

* CE IVD kits only available in Germany
** Release planned for 31st March 2020

3. Amplification and Detection

Molecular assays to detect SARS-CoV-2 have been developed and are accessible through the homepage of the WHO. Furthermore, several commercial kits based on Real-time PCR are available such as:

• Coronavirus (Strain 2019-nCoV) by Genesig/Primerdesign
• SARS-CoV-2 detection assay by TIB MOLBIO
• RIDA® GENE Novel Coronavirus 2019 RUO Tests by r-biopharm

The PCR set up can be pipetted by a liquid handling platform like CyBio FeliX . Using PCR workstations or cabinets increases maximum safety and minimizes contamination risks. Single components such as reverse transcriptase or others are available via Analytik Jena as well. In general all Real-time PCR-based assays can be established on Real-time thermal cycler of the qTOWER³ family provided by Analytik Jena or similar.
https://www.who.int/emergencies/diseases/novel-coronavirus-2019/technical-guidance/ laboratory-guidance