A Specific miRNA Encoded by SARS-CoV-2 as a Diagnostic Tool to Predict Disease Severity in COVID-19 Patients
Condition: COVID-19
MeSH terms: COVID-19
Study type: Interventional

Summary: 
      Coronavirus disease is caused by SARS-CoV-2, known as 2019 novel coronavirus (2019-nCoV). To
      date has caused a large number of deaths causing serious respiratory illness such as
      pneumonia and lung failure, therefore representing a serious threat to public health. The
      etiological agent belongs to the subfamily Orthocoronavirinae in the family Coronaviridae,
      Order Nidovirales. The genome of coronaviruses is composed of an enveloped, positive-sense,
      single-stranded RNA with a size varying between 26 kb and 32 kb, becoming the largest genome
      of known RNA viruses so far. Similar to RNA viruses, this family is characterized by genetic
      variability and high recombination rate that enable them to be easily distributed among
      humans and animals worldwide. Considering the huge impact of the pandemic, it is urgent to
      gain understanding and to build strategies to contain the viral spread. To date, different
      diagnostic kits for testing the illness are available. Besides diagnosis, the prediction of
      the severity and prognosis of COVID-19 is essential to stratify patients and allocate them in
      the adequate medical facilities so as to reduce mortality rates. It has been reported that
      microRNAs (miRNAs) are valuable biomarkers for disease diagnosis, prognosis and
      classification. MiRNAs are defined as a class of non-coding RNAs that are able to regulate
      gene expression by specific binding to complementary regions in coding messenger RNAs,
      leading to translational repression or decay. Not only that, but also they can be important
      modulators of viral infections.Previous studies have revealed the presence of miRNA-like
      small RNAs (milRNAs), which can be encoded by RNA viruses and can actively disrupt the host
      innate immune responses in order to create a favourable environment for viral replication. On
      the other side, cellular miRNAs can also play a role on virus replication and pathogenesis.In
      this case, this pilot project is aimed at their valuable diagnostic potential, in order to
      diagnose and stratify patients under viral infection. The project came up after receiving
      information from a Chinese research group, requesting their results to be replicated in
      Caucasian population. The ROC curves were constructed to demonstrate the accuracy of this
      specific miRNA in COVID-19 patient stratification and discerning between severe patients from
      healthy controls. Both ROC curves suggested the miRNA as precise biomarker for differential
      diagnosis and prognosis of disease severity.
    

Detailed description: 
      It will include 120 patients with and without COVID-19 According to the World Health
      Organization guidance, all patients with COVID-19 enrolled in the study were diagnosed bythe
      throat swab and RT-PCR method. Patient information was collected from medical records.Total
      RNA content will be extracted from plasma samples by methods aimed at preserving and
      isolating small RNA molecules. The kit used will be. mirVANATM PARISTM Isolation Kit (Applied
      Biosystems, Darmstadt, Germany). The mirVANATM PARISTM Kit begins with homogenization of
      samples with a special cell disruption buffer and RNA isolation is performed using a
      procedure that combines the advantages of organic and solid-phase extraction. Once extracted,
      a fixed volume of RNA solution will be used as the input for the reverse transcription
      reaction using the TaqMan™ MicroRNA Reverse Transcription Kit (Applied Biosystems, Darmstadt,
      Germany). The product will be further amplified by real-time PCR using specific primers for
      the detection of the milRNA previously validated by means of the LightCycler® 480 Real-Time
      PCR System (Roche Diagnostics, Barcelona, Spain).
    

Intervention name(s): miRNA analysis in plasma
Intervention type(s): Diagnostic Test
Intervention description(s): blood sample
Allocation: Non-Randomized
Intervention model: Parallel Assignment
Masking: None (Open Label)
Primary purpose: Screening