Presented by: Manuela Álvarez Villegas Tercer semestre Curso de Biología Molecular Universidad Pontificia Bolivariana

1. Presented by:
Manuela Álvarez Villegas
III semestre

2. INTRODUCTION
ZIKA VIRUS HEPARAN SULFATE
✓ Is an emerging arbovirus of the Flaviviridae
family.
✓ The infection of ZIKV often leads to self-limiting
diseases.
✓ It causes severe neurologic disorders including
microcephaly and Guillain–Barré syndrome.
✓ no specific vaccine and drug is available for the
treatment of these ZIKV diseases.
✓ It´s a linear polysaccharide found in almost all
animal tissues.
✓ It has been identified as an attachment factor for
several flaviviruses.
✓ HS is associated with proteins of cell surface and
from extracellular matrix.
✓ HS is employed as a nonspecific attachment
factor by a number of viruses.

3. Zika virus and heparan sulfate
Tan et al
reported that the ZIKV virus particles do
not bind to heparin, a glycosaminoglycan that has similar
protein binding properties to HS
Kim et al
showed that the recombinant ZIKV envelope protein
interacts with various glycosaminoglycans including
heparin
➢ ZIKV did not rely on the HS for its entry.
➢ The early replication of ZIKV was reduced in the HS
deficient cells
➢ HS had an impact on the ZIKV-induced apoptosis.

4. OVERALL OBJECTIVE
• Study the role of heparan
sulfate in the Zika virus entry,
replication, and cell death

5. MÉTODOS
PCR RTQ
ARNm
Transcriptasa inversa
ADNc
Permite cuantificar las secuencias específicas
de ácidos nucleicos mediante el uso de
reporteros fluorescentes en la reacción.
➢ Utiliza sondas marcadas con un marcador fluorescente
➢ Durante el ciclo, la sonda de desnaturaliza y se acopla a la secuencia objetivo
➢ La PCR RTQ utiliza dos instrumentos principales:
1. Termociclador : desnaturalización, hibridación y elongación.
2. Módulo óptico: Mide la intensidad de la fluorescencia.
→ Cuando se miden 30 ciclos se convierte en una curva de amplificación

6. CITOMETRÍA DE FLUJO Permite medir diferentes parámetros de una célula
➢ Se basa en hacer pasar una suspensión de partículas
alineadas por un haz luminoso.
➢ Esto produce señales luminosas que son detectadas y
transformadas a señales eléctricas
➢ Estas se amplifican y se convierten en
señales digitales que son procesadas por un
computador.

7. MTT Compuesto AMARILLO soluble en agua.
✓ Cuando se reduce por medio de la succinato
deshidrogenasa, se convierte en un compuesto VIOLETA
insoluble en agua.
Disolvente orgánico
Cuantificación del
color
➢ Funcionabilidad
mitocondrial de las
células
➢ Mide:
1. Supervivencia
2. Proliferación celular

8. WESTERN BLOT
Es usado para detectar
proteínas especificas en
una muestra determinada.
✓ proteína ZIKV E
✓ proteína DENV2
1. Se separan las proteínas mediante una electroforesis en gel, según
el parámetro que se desee:
- Peso molecular
- Estructura
- Hidrofobicidad
2. Se transfieren a una membrana de nitrocelulosa
3. Se busca la proteína de interés con anticuerpos específicos para ella
4. Se detecta la interacción antígeno anticuerpo

9. RESULTADOS

12. DISCUSSION
AUTOR WHAT THEY SAID OR DID YES NO
Hamel et al
Although ZIKV shares many cell
surface molecules such as DC-SIGN,
and AXL with DENV their
requirements on HS are differential
Tan et al
showed that the majority of ZIKV
viral particles were not retained by
heparin sepharose, and the
inhibition of cell sulfation by sodium
chlorate or the removal of HS by
heparinase I/III did not affect viral
infectivity, suggesting the
virus does not bind HS.
Chen et al
Heparan sulfate has been identified
to be an attachment factor for
several flaviviruses, including DENV,
YFV, and JEV

13. CONCLUSIONS
1. In this study, there is no complete clarity on whether heparan
sulfate plays a role in the infection of the virus, but its future
knowledge will allow designing methods that can intervene and
prevent the replication of the virus.
2. The timely diagnosis of Zika is vital to avoid complications that
further deteriorate the patient´s health status.

15. GRACIAS!