This study aimed to detect the SARS-COV2 viral component directly from inoculated VTM without RNA extraction. Inoculated VTMs of already tested 50 positive and 50 negative samples were divided into three groups. Group I was treated with Proteinase K (PK) followed by 3-step-heat treatment at different temperatures (25°C, 60°C, and 98°C) and stored at 4°C. Group II was directly subjected to 3-step-heat treatment without PK exposure and stored at 4°C. And group III was set-up as standard group; it was processed using Qiagen’s column based QIAamp Nucleic Acid kit and the obtained nucleic acids were stored at 4°C. These stored samples were used as a template to execute real-time polymerase chain reaction, and results were noted. Group I demonstrated 96% and 88% sensitivity for N and ORF1ab genes respectively, whereas group II demonstrated 78% and 60% when compared to the results of standard group III. Overall group I showed better results than group II when compared to group III. Thus, in situations where gold-standard reagents are not available, PK exposure and heat treatment can be employed to carry out molecular detection of SARS-CoV2 viral component.
【저자키워드】 SARS-CoV-2 detection, heat inactivation, real time PCR, COVID – 19, Proteinase K, 【초록키워드】 Treatment, SARS-CoV2, RNA extraction, polymerase chain reaction, nucleic acid, sensitivity, Viral, VTM, nucleic acids, real-time polymerase chain reaction, temperature, molecular, group, Proteinase K, ORF1ab gene, followed by, Chain Reaction, Template, three groups, acid, positive, ORF1, reagent, negative sample, polymerase chain, tested, detect, inoculated, treated, were used, demonstrated, processed, 【제목키워드】 SARS-CoV2, detection, RNA, extraction, vitality,