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Genotypic Characterization of African Swine Fever
Virus Isolated from Outbreaks in Kenya between
2011 and 2013
Cynthia Onzere
MSc. International Animal Health
University Of Edinburgh
18th October 2013
Background & Significance
The study explores four genetic markers in addition to the B646L gene to identify genotypes and
determine variations within and between genotypes. The study also intends to eventually evaluate the
effects of the variations on the pathogenicity of the virus. These markers include:
The complete E183L gene that encodes the p54 ASFV protein
essential in the recruitment of envelope precursors to the
assembly site (Rodriguez et al., 2004).
The variable 3′-end of the B646L gene that encodes the
major capsid protein p72
Inner membrane
Matrix
The B602L gene that encodes the central variable region (CVR)
where repeated amino acid tetramers that vary in number and
type among ASFV isolates are located. This variation is
important in identifying and grouping the ASFV isolates.
Red Blood cell
Infected leucocyte
The EP402R gene encodes
the CD2v protein that is
responsible for erythrocytes
haemadsorption around
ASFV infected cells (Borca et
al.,1998).
The CP204L gene that encodes
the p30 protein that Modifies
the subcellular distribution of
heterogeneous nuclear
ribonucleoprotein K and may
contribute to modulate
HNRNPK functions related to
processing and export of
mRNAs during ASFV infection.
Conceptual or theoretical framework
Materials and Methods
ASFV diagnosis andverification usingconventional PCR,UPL and TCORPCR and selectionof ASFV positivesamples.
Collection of ASFV naïve blood for PBMC isolation. Culture of the leucocytes using RPMI medium and autologous serum and infection of the resultant macrophages with the ASFV isolates.
Monitoring haemadsorption (HAD)
Harvesting and extraction of DNA from HAD positive cultures.
Genotyping and sequencing of thepartial and full length VP72, VP54,and CVR markers.
Nucleotide and molecular evolutionary analyses using CLC workbench, MEGA version 5.2, Mobyle and Bioedit
Blood, tissues and serum samples are obtained from the ASF cross sectional survey, longitudinal survey and suspected outbreak areas
Purification
P72
CVR
ken12/bus.sig
ug13.231
ken12/burumba.F1
P73
P72
P69
ken12/kia.p59
P43
ken12/nrb.p39
ken12/nrb.p38
ken12/nrb.P36
ken12/kia.p32
ken13/kiri.P31
ken13/kia.P30
ken12/athi.p26
ken12/athi.p24
P21
P09
ken13/bus.obai
Ken13/bus.1033
ken13/bus.564
ken13/bus.125
ken11/kisauni.P52
ken11/Kisauni.p13
Ug12.Kabale1
Ug10.Adjumani
Ug11.Mpigi
Ug10.Moyo2
Ug12.Lira
Ug12.Kampala3
Ug13.Busia2
UGA/1/95
UG07.Wak4
Ken11/Bus1.2
Ken11/Kia2.1
Ken11/ThikP06
Ken11/KakSP
Ken10/KAKFA1
Ken10/Kis027
Ken06.Bus
Ken06.Kis
Ken07.Eld2
Ken07.Kia
Ken07.Nak
Ken08WH/8
Ken05/Tk1
Ken05/Tk6
Ken05/Tk10
Ken05.DPN2
Ken09Tk.15/6
Ken08Tk.2/1
Ken09Tk.15/4
Ken09Tk.19/7
57
62
61
69
98
0.002
P54
IX
X
Dendrograms generated using three marker loci
ken12/nrb.p39
ken12/kia.p59
ken12/nrb.p38
ken12/nrb.P36
ken12/kia.p32
ken13/kiri.P31
ken13/kia.P30
ken12/athi.p26
ken12/athi.p24
ken13/bus.125
ken12/burumba.F1
ken11/kisauni.P52
ken11/Kisauni.p13
Ug10.Kumi
Ug13.Busia2
Ug12.Kyenjojo
Ug03P.6
Ken06.Kis
Ken07.Nak
Ken08WH/4
UG07.Wak4
Ken10/Kis027
Ken11/Bus1.2
Ken11/ThikP06
Ug12.Lira
Ug11.Mpigi
Ug10.Adjumani
Ug12.Kabale1
Uga 95/1
86
64
0.005
ug13.231
ken12/athi.p26
ken12/kia.p59
ken12/nrb.p38
ken12/kia.p32
ken12/nrb.P36
ken13/bus.125
ken11/kisauni.P52
ken11/Kisauni.p13
Ug12.Kabale1
Ug10.Kumi
Ug10.Adjumani
Ug11.Mpigi
Ug12.Lira
Ug13.Busia2
UG07.Wak4
Ken11/Bus1.2
Ken11/Kia2.1
Ken11/ThikP06
Ken10/KAKFA1
Ken10/Kis027
Ken08WH/4
Ken06.Kis
Ken07.Nak
ken12/athi.p24
UGA/95/1
0.002
THANK YOU
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