This document summarizes a study that detected West Nile Virus lineage 2 in Culex pipiens mosquitoes in Greece in 2017. The study used PCR and RT-PCR techniques to extract genomic DNA and RNA from mosquitoes to analyze biotypes and detect the presence of flaviviruses like West Nile Virus. Lineage 1 and lineage 2 of West Nile Virus were able to be simultaneously detected. The results and discussion sections indicate the virus was found in local mosquitoes, and that Culex pipiens is likely the main vector of West Nile Virus transmission in the region due to its behavior of biting both birds and humans. While West Nile Virus has many vectors, correct prevent

1. Karoll Lucía León T
III Semester
Molecular Biology
UPB - 2018 - II

2. INTRODUCTION
WEST NILE VIRUS
Single-stranded RNA
molecule of positive
polarity (11 kb)
3 structural proteins:
capsid, envelope, and
premembrane.
7 nonstructural
proteins: NS1, NS2A,
NS2B, NS3, NS4A,
NS4B, and NS5.

3. INTRODUCTION
CULEX PIPIENS MOSQUITO
• The complete life cycle of
mosquitoes takes about 13–15
days to complete.
• It is considered as the main
vector of several viruses and
diseases.
• Is normally considered to be a
bird feeder but some urban
strains have a predilection for
mammalian hosts and feed
readily on humans.

4. GENERAL OBJECTIVE
Detection of West Nile
Virus – Lineage 2 in Culex
pipiens mosquitoes,
associated with disease
outbreak in Greece,
2017.

5. MATERIALES Y MÉTODOS
•PCR
FUNDAMENTO: Amplificar una
secuencia de material genético
(ADN).
Se extrajo ADN genómico
(ADNg) de cada uno de los
mosquitos y se analizó el
biotipo con el uso de un ensayo
diagnóstico de PCR.

6. MATERIALES Y MÉTODOS
•PCR
La presencia y frecuencia de la
mutación kdr en el sitio L1014F
en el canal de sodio controlado
por voltaje se detectó con un
ensayo de PCR específico de
alelo.
Se detectó, con una reacción de
PCR, la presencia de mutaciones
en el extremo C-terminal de la
quitina sintasa que abarcaba la
posición 1043.

7. MATERIALES Y MÉTODOS
•RT – PCR
FUNDAMENTO: Amplificar
una secuencia de material
genético (ARN)
Se realizó una RT-PCR de un
paso para evaluar la
presencia de Flaviviruses

8. MATERIALES Y MÉTODOS
•RT - PCR
La detección de WNV se
realizó mediante un ensayo
multiplex de TaqMan de RT-
PCR en tiempo real de un
solo paso.
Detecta y diferencia,
simultáneamente, el WNV-
linaje 1 y el WNV-linaje 2.

9. RESULTADOS

10. RESULTADOS

11. DISCUSSION
AUTHOR WHAT DID HE/SHE SAY? YES OR NO
Hernandez-Triana et al., 2014;
Chaskopoulou et al., 2011
Virulent and neuroinvasive strains do
exist in Europe and Greece with similar
ratios of infection reported for lineage 1,
leading to clinical disease.
Patsoula et al., 2016 The high abundance of Cx. pipiens
mosquitoes in the area and the detection
of WNV implies these mosquito species
were most likely the main WNV vector in
the region, as is the case in Northern and
Central Greece.
Ciota et al., 2013 Cx. pipiens/molestus are considered an
extremely competent vector of WNV due
to their enhanced WNV transmission
potential.

12. CONCLUSIONS
The main vector of the WNV is the
Cx pipiens pipiens/molestus due
to its behavior. This mosquito
bites birds which causes the
maintining of the virus among this
hosts and facilitate its
transmission to humans.
Despite having a large number of
WNV vectors in this region, with
the correct measures of
prevention and control the virus
should not manifest itself in
many individuals.