Kit is used for detecting the epidemic virus “SARS-CoV-2 (2019-nCoV)” causing Coronavirus Disease 2019 (COVID-19). The kit is applied to nucleic acid isolates from nasopharyngeal aspirate / lavage, bronchoalveolar lavage, nasopharyngeal swab, oropharyngeal swab and sputum samples. Rapid diagnosis with the kit is achieved via one-step reverse transcription (RT) and real-time PCR (qPCR) (RT-qPCR) targeting SARS-CoV-2 (2019-nCoV)-specific RdRp (RNA-dependent RNA polymerase) gene fragment. The RdRp gene-targeted Wuhan-RdRp oligonucleotide set gives positive results only with SARS-CoV-2 (2019-nCoV). In the SARS-CoV-2 (2019-nCoV) routine screening,
Wuhan-RdRp olionucleotide set is applied; if the result is Wuhan-RdRp positive, SARS-CoV-2 (2019-nCoV) is interpreted as positive; and if the result is Wuhan-RdRp negative SARS-CoV-2 (2019-nCoV) is interpreted as negative.
The kit is validated with RINA TM M14 Nucleic Acid Extraction Robot (Cat No: RINA-M14-01) and its consumables (Cat No: RN-NA-14-111-100) and the manual extraction kits BS-NA-109 (respiratory tract samples) and BS-NA-102-100 (all the sample types except feces). The kit is validated for 10 and 20 µL qPCR volumes using Roche LightCycler® 96, Bio-Rad CFX96 Touch™, Qiagen RotorGene® 5 Plex Real-Time PCR Systems. The kit’s detection limit (LOD) is the lowest analyte concentration that can be detected with a 95% probability. The LOD of the oligonucleotide sets included in the kit, inclusivity and exclusivity studies were carried out using 35 different genotypes and published by WHO (WHO Protocol, 13.01.2020, Diagnostic detection of Wuhan coronavirus 2019 by real-time RT-PCR). Analytical and clinical performance of the kit was determined by the “Turkish Ministry of Health, General Directorate of Public Health, Department of
Microbiology Reference Laboratories and Biological Products (HSGM)”. HSGM tested the inclusivity with 38 different clinical samples confirmed by DNA sequence analysis, and the exclusivity with Coronavirus 229E/OC43/NL63/HKU1, Influenza A H1/H3, Influenza B, Parainfluenza 1/2/3/4, Metapneumovirus, Rhinovirus, Respiratory syncytial virus (RSV) A/B, Bocavirus (BoV), Enterovirus, Adenovirus, Legionella pneumophila, Haemophilus influenzae, Streptococcus pneumoniae, Bordetella pertussis. The LOD of the kit determined by HSGM is 5.6 copies SARS-CoV-2 genome/reaction. The kit does not cross-react with other respiratory pathogens. The kit was applied concurrently with DNA sequence analysis to SARS-CoV-2 positive 500, SARS-CoV-2 negative 500 clinical samples. The analytical sensitivity of the kit is 99.4% and its specificity (specificity) is 99.0%.