| Grant number: | 18/23680-0 |
| Support Opportunities: | Scholarships in Brazil - Master |
| Start date: | April 01, 2019 |
| End date: | November 30, 2020 |
| Field of knowledge: | Biological Sciences - Microbiology - Applied Microbiology |
| Principal Investigator: | Edison Luiz Durigon |
| Grantee: | Camila Pereira Soares |
| Host Institution: | Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil |
Abstract RSV is a virus widely distributed in the world and is the main virus involved in lower respiratory tract infections. The population at risk for these infections are children under 5 years of age, immunocompromised patients and the elderly. The RSV belongs to the family Pneumoviridae, its genome is composed of a single stranded RNA molecule and encodes 11 proteins. Divided into 2 antigenic groups based on G (A and B) genotyping, it has several well characterized genotypes within each group. The similarity between the symptoms caused by RSV infection with other respiratory viruses impairs the accurate clinical diagnosis. In this context, laboratory diagnostic methods are extremely important. Firstly, because they prevent the common practice of antibiotic prescribing and because patient isolation measures can be applied by decreasing the degree of viral spread. Some novelties have arisen in recent years regarding molecular diagnosis, however, one has been presented in a very promising way: the CRISPR-Cas technique. The technique that uses the Cas9 bacterial endonuclease to edit DNA sequences has already been widely used, and now new proteins have started competing in the spotlight. Two articles published in early 2018 showed the success of two techniques that used proteins like Cas13 and Cas12 in the diagnosis of viruses such as Zika and HPV. The SHERLOCK technique (Specific High-Sensitivity Enzymatic Reporter Unlocking) was able to detect high sensitivity and specificity of viral RNA samples in a few minutes using the Cas13a protein. A characteristic of this technique that makes it simple and fast is the use of the RPA (Reaction Polymerase Amplification) technique as a method of amplification of the genetic material that, unlike PCR, is isothermal, and does not require the use of thermocycling devices. Our objective in this work is to standardize the SHERLOCK technique, using the Leptrotricia wadeii Cas13a protein to detect Human Respiratory Syncytial Virus in samples of children with acute respiratory disease and compare with the standard technique of Real Time PCR | |
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