Screening and Detection SARS-CoV-2RNA from Buffy Coats v1

This SOP has been developed to minimize or eliminate employee occupational exposure to SARS-CoV-2 virus from blood samples of healthy volunteers obtained from the New York Center (NYBC) or from outpatients at HSS. Both are screened for symptoms and potential exposure at the point of care but are not tested for COVID-19. The CDC 2019-Novel Coronavirus (2019-nCoV) Real-Time RT-PCR Diagnostic Panel is intended for the qualitative detection of nucleic acid from the 2019-nCoV in upper and lower respiratory specimens (such as nasopharyngeal or oropharyngeal swabs, sputum, lower respiratory tract aspirates, bronchoalveolar lavage, and nasopharyngeal wash/aspirate or nasal aspirate). This protocol is a modified version of the CDC protcol meant to screen for Novel Coronavirus (2019-nCoV) in RNA isolated from whole blood. The Real-Time RT-PCR is for the identification of 2019-nCoV RNA from buffy coats obtained from supposedly healthy donors from New York Blood Center (NYBC). The 2019-nCoV RNA is generally detectable in upper and lower respiratory specimens during infection while rarely detected in circulating blood. Positive results are indicative of active infection with 2019-nCoV and blood should be discarded as per the . Negative results do not preclude 2019-nCoV infection and should not be used as the sole basis of management decisions. The product contains oligonucleotide primers and dual-labeled hydrolysis probes (TaqMan®) and control material used in rRT-PCR for in vitro qualitative detection of 2019-nCoV RNA. The panel is designed for specific detection of the 2019-nCoV (two primer/probe sets). An additional primer/probe set to detect the human RNase P gene (RP) in control samples. RNA is purified from whole blood in one-step Real-Time RT-PCR on an Applied Biosystems 7500 Real-Time PCR Instrument. Detection of viral RNA in whole blood aids in the safe operation of research labs working with human blood samples.