Validation of Laboratory Techniques, Strategies, and Types of Samples for Epidemiological Control in the Covid-19 Pandemic

• RT-PCR (Diagnostic test), also known as Real-Time Polymerase Chain Reaction: RT-PCR (Gold standard): Real-time PCR testing with reverse transcriptase (RT-PCR). The test consists of a polymerase chain reaction that has previously had a reverse transcription phase, RT. This test allows the cADN (complementary DNA) to be obtained from a RNA chain. This technique allows the detection and amplification of a sequence from a strand of RNA.
• LAMP (Diagnostic test), also known as Loop-mediated isothermal amplification: LAMP: The LAMP technique, like PCR, has the ability to amplify specific DNA fragments (target sequence), allowing for highly sensitive detection of pathogens. Unlike pcr, LAMP requires between 4 and 6 feeders, external F3 and B3 feeders, which hybridte with the outer regions of the target sequence, while the internal FIP and BIP have sequences in both ways that allow the formation of a loop. The LAMP reaction is isothermal, i.e. it takes place at a single temperature. LAMP has been successfully used to detect viral diseases as well as viral pathogens such as West Nile virus, TMR-1, Norovirus, avian influenza, foot-and-mouth disease, classical swine fever, among others.
• POOL RT-PCR (Diagnostic test), also known as Pooled Real-Time Polymerase Chain Reaction: POOL RT-PCR: This test aims to mix in a single sample the extractions obtained from several people. The joint sample is evaluated and, if negative, it is understood that all are free of COVID-19; if it is positive, an array system is used to detect the affected person. That is, a single PCR (polymerase chain reaction), the diagnostic test used to detect the virus pathogen is applied to a group and repeated individually if positive.
• POOL LAMP (Diagnostic test), also known as Pooled Loop-mediated isothermal amplification: POOL LAMP: Like the RT-PCR pool test. This test essentially involves mixing in a single sample the extractions obtained from several people. This joint sample is tested and, if it is negative, it is understood that everyone is free of COVID-19; if it is positive, an array system is used to detect the affected person. That is: a single LAMP applies to a group

Primary outcomes

• Validation [1 month]: Validate different molecular tests and sampling and sample type strategies used in epidemiological control of COVID-19 diagnosis in Tarija, Bolivia.

Secondary outcomes

• RT- PCR (Saliva) [1 week]: Validate the use of molecular testing based on the PCR method using Saliva samples
• LAMP [1 week]: Validate the use of the molecular test based on the LAMP method using saliva and nasopharyngeal samples
• POOL PCR [1 week]: Validate the use of molecular tests with Pool PCR strategy using saliva samples
• POOL LAMP [1 week]: Validate the use of molecular tests with Pool LAMP strategy using saliva samples

Sex: All, ages: 21 Years to 64 Years, accepts healthy volunteers

Inclusion criteria:

* Symptomatic: Subjects with signs and symptoms of respiratory infection less than or equal to 3 days, preferably with clinical and molecular diagnosis compatible with Covid-19.
* Asymptomatics: Subjects who have had direct contact with people infected and who have not shown any symptoms related to Covid-19.
* Negative: Individuals with negative RT-PCR testing for SARS-CoV-2 (reference test) who have not manifested any symptoms seven days prior to sampling.

Exclusion criteria:

* Adults under the age of 21 or over 65
* Subjects with a clinical history of autoimmune disease or chemotherapy treatments.
* Subjects who have received transfusions or convalescent plasma in the last month prior to enrollment.
* Subjects with prior diagnosis of other infectious diseases such as, but not limited to AIDS, Hepatitis B or C, Tuberculosis, or any other ongoing or unresolved diseases.
* Subjects with malformation or oncological pathology of the upper respiratory track that may hinder sample collection, including but not limited to deviated septum, allergic rhinitis.
* Subjects who have not signed or can not sign the informed consent form
* Subjects not able to provide their complete information or access to their clinical history.

• Kenji Shoji, PhD, Principal investigator, Lakefront Biotherapeutics NV
• Rodrigo Arce Cardozo, MD, MPH, Study chair, City University of New York, School of Public Health