Performance verification (secondary validation) of the real-time molecular PCR (RT-qPCR) test on the Vivalytic One®* (Vivalytic SARS-CoV-2) platform for point-of-care (POC) detection of SARS-CoV-2.

The new SARS-CoV-2 coronavirus (severe acute respiratory syndrome, coronavirus type 2) is an encapsidated single-stranded RNA-positive virus easily transmitted to humans causing COVID-19 disease (1). This virus is a member of the Coronaviridae family and belongs to the Betacoronavirus genus along with HCoV-OC43, HCoV-HKU1, and the highly pathogenic viruses SARS-HCoV (severe acute respiratory syndrome) and MERS-HCoV (Middle East respiratory syndrome) (1).
On January 30, 2020, the World Health Organization (WHO) declared the SARS-CoV-2 virus a public health emergency of international concern after it rapidly spread worldwide causing the COVID-19 pandemic (1). The first reports of positive cases for this virus were presented in December 2019 in Wuhan, China and since that date more than 84 million confirmed cases and more than one million eight hundred thousand deaths have been reported worldwide, with the majority of cases found in the American continent (WHO daily report, https://covid19.who.int/).
In the particular case of Colombia, according to the report of the National Health Institute, to date more than one million six hundred thousand confirmed cases and more than forty-four thousand deaths have been reported (https://www.ins.gov.co/Noticias/paginas/coronavirus.aspx). Taking into account these figures and in order to promote clinical and public health interventions to counteract this health emergency, it is necessary to carry out an extensive processing of samples for a prompt and adequate in vitro diagnosis of SARS-CoV-2. To this end, it is important to identify and validate protocols that can be implemented in a rapid, automated, low-cost manner with good sensitivity and specificity results.
One of the first validated protocols for the diagnosis of SARS-CoV-2 was the protocol developed by the Charité Virology hospital in Berlin, Germany (2). This protocol, aimed at detecting the E (Envelope gene) and RdRp (RNA-dependent RNA polymerase gene) genes by means of three consecutive real-time Polymerase Chain Reaction (PCR) reactions, has now been implemented in laboratories around the world and is recommended by the WHO (2). In Colombia, it is used as the reference standard for the verification of several tests developed for the detection of SARS-CoV-2 (3).
Timeliness in the diagnosis of SARS-CoV-2 infection is a recognized need for clinical decision making, preventive isolation and potentially reducing transmission (3). In view of this, rapid processing devices have been developed, where sample handling is minimized and results are generated at the patient's site of care in less than two hours. These tests are known as "Point-of-care tests" (POCT). In Colombia, POCTs must undergo a verification process (secondary validation) to determine that the performance conditions published by the manufacturer meet the quality standards of the Colombian Ministry of Health and Social Protection (4).
However, many of the POCT tests use microfluidic systems whose performance can be affected by environmental conditions, especially atmospheric pressure conditions. This requires performance verification under the conditions in which it will be used, specifically at altitudes above 2500 meters above sea level, an atmospheric condition that applies to a significant number of cities in Colombia and South America.
Taking into account the above mentioned, this document proposes the validation of this protocol under the atmospheric pressure conditions of Bogota.

Molecular Test, PCR, SARS-CoV-2