Focus Simplexa Controls
The DiaSorin Molecular Simplexa™ COVID-19 Direct real-time RT-PCR assay is intended for use on the LIAISON® MDX instrument for the in vitro qualitative detection of nucleic acid from severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in nasopharyngeal swabs (NPS), nasal swabs (NS), nasal wash/aspirate (NW) or bronchoalveolar lavage (BAL) specimens from individuals suspected of COVID-19 by their healthcare provider. The Simplexa COVID-19 Direct assay is an aid in the diagnosis of SARS-CoV-2 infection. Testing is limited to laboratories certified under the Clinical Laboratory Improvement Amendments of 1988 (CLIA), 42 U.S.C. §263a, to perform high or moderate complexity tests.
Results are for the identification of SARS-CoV-2 RNA. The SARS-CoV-2 RNA is generally detectable in upper respiratory and
bronchoalveolar lavage (BAL) specimens during the acute phase of infection. Positive results are indicative of the presence of
SARS-CoV-2 RNA; clinical correlation with patient history and other diagnostic information is necessary to determine patient
infection status. Positive results do not rule out bacterial infection or co-infection with other viruses.
The agent detected may not be the definite cause of disease. Laboratories within the United States and its territories are required to report all positive results to the appropriate public health authorities. Negative results do not preclude SARS-CoV-2 infection and should not be used as the sole basis for patient management decisions. Negative results must be combined with clinical observations, patient history, and epidemiological information.
The Simplexa™ COVID-19 Direct assay is intended for use by qualified and trained clinical laboratory personnel specifically
instructed and trained in the techniques of real-time PCR and in vitro diagnostic procedures. Simplexa™ COVID-19 Direct is only for use under the Food and Drug Administration’s Emergency Use Authorization.
SUMMARY AND EXPLANATION
SARS-CoV-2 (also called COVID-19 virus) is a beta coronavirus belonging to the family of Coronaviruses, named for the crown- like spikes on their surface. There are four main sub-groupings of coronaviruses, known as alpha, beta, gamma, and delta. Common human coronaviruses are 229E (alpha coronavirus), NL63 (alpha coronavirus), OC43 (beta coronavirus) and HKU1 (beta coronavirus), and these usually cause mild to moderate upper-respiratory tract illnesses, like the common cold.1,2,3 Other human coronaviruses such as MERS-CoV (the beta coronavirus that causes Middle East Respiratory Syndrome, or MERS) and SARS-CoV (the beta coronavirus that causes severe acute respiratory syndrome, or SARS) have caused more severe respiratory illness with higher rates of morbidity and mortality.
The SARS-CoV-2 is a novel coronavirus that causes coronavirus disease 2019, or COVID19. SARS-CoV-2 caused an outbreak beginning in December 2019 in Wuhan City, Hubei Province, China and has spread globally, being consequently declared a pandemic by the World Health Organization (WHO).2,4 Patients with COVID-19 have had mild to severe respiratory illness with symptoms of fever, cough and shortness of breath, and many patients have had complications including pneumonia in both lungs.5
PRINCIPLES OF THE PROCEDURE
The DiaSorin Molecular Simplexa™ COVID-19 Direct assay system is a real-time RT-PCR system that enables the direct
amplification of Coronavirus SARS-CoV-2 RNA from nasopharyngeal swabs (NPS), nasal swabs (NS), nasal wash/aspirate (NW) or bronchoalveolar lavage (BAL) specimens. The system consists of the Simplexa™ COVID-19 Direct assay, the LIAISON® MDX (with LIAISON® MDX Studio Software), the Direct Amplification Disc and associated accessories.
In the Simplexa™ COVID-19 Direct assay, fluorescent probes are used together with corresponding forward and reverse primers to amplify SARS-CoV-2 viral RNA and internal control RNA. The assay targets two different regions of the SARS-CoV-2 genome, ORF1ab and S gene.
The S gene encodes the spike glycoprotein of the SARS-CoV-2 (COVID-19 virus) and is also targeted to
specifically detect the presence of SARS-CoV-2. The ORF1ab region encodes well-conserved non-structural proteins and therefore is less susceptible to recombination. An RNA internal control is used to detect RT-PCR failure and/or inhibition.
The Simplexa™ COVID-19 Direct assay contains sufficient reagents for 24 reactions. Upon receipt, store at -10 to -30 ºC (do not use a frost-free freezer). Each vial contains sufficient material for one use. Use within thirty (30) minutes ofthawing.