The Relationship between Factor V Leiden
Mutation and Recurrent Abortion Among
Palestinian Pregnant Women
In The West Bank
Ayman Hussein
An-Najah National University
1
Recurrent Pregnancy loss
 Defined as 3 or more spontaneous fetal
losses before the 20th week of pregnancy .
 A major health problem affecting 0.5%1.0% of pregnant women
 occurring in about 20 % of all pregnancies,
fewer than 50% of these cases have
definitive causes .
2
Thrombophilia
• It is a predisposition to thrombosis.
• It is a multigenic disorder caused by
inherited and acquired factors
• it contributes to the increasing incidence
of poor pregnancy outcomes including
recurrent abortion .
3
Factor V
 Factor V protein 330 k Da made chiefly by
cells in the liver ,circulates in plasma as a
single-chain molecule with a plasma halflife of about 12 hours1984).
 In contrast to most other coagulation
factors, it is not enzymatically active but
functions as a cofactor
4
 Pregnant women have 2–5-fold higher risk for
venous thromboembolism (VTE )compared to
non-pregnant women.
 The FVL variant allele increases the risk for
(VTE) during pregnancy 8–10 times,
compared to non-pregnant women with the
wild-type genotype .
5
Factor V
Procoagulant and Anticoagulant Actions of Factor V.
 Procoagulant : , factor
Va enhances the
activation of
prothrombin (factor II)
by factor Xa.
 Anticoagulant :Factor V
can serve (with protein
S) as a cofactor in the
inactivation of factor
VIIIa by activated
protein C (APC)
Group
Total No
NO
%
Of
OF
women
mutatio
Mutation genotype%
Comparison with the control
group
homo
hetro
P value
0%
22%
0.594
n
Control
50
11
group
Primary
(22 %)
32/54
8
y
ratio
2.787
CI %95
(2.1575,
3.4165)
(25.0
6.30%
18.70%
0%
31.80%
%)
abortion
Secondar
Odds
7
22/54
31.8%
abortion
6
Leiden Mutation
Single Point
Arginine
4/13/2015
‫خالد شلباية‬
R506Q
Glutamine
7
Factor V Leiden mutation
 Inheritance is autosomal dominant.
Heterozygous :4.5-14.5%
Homozygous : 1:5000
 Present in 4 to 10% of people of Caucasian origin
(Bertina et al 1994; Rees 1996).
 The most common cause of inherited thrombophilia,
accounting for 20% to 50% of cases (Bertina et al
1994).
 Both homo. and heterozygous individuals with
the mutation can develop thrombosis. Homo have
91 times increase in thrombosis .
8
Objective
The main objective of our study was :
To investigate if there is a significant
association between factor V Leiden
mutation and Recurrent Abortion among
Palestinian Pregnant Women In the West
Bank.
9
Methodology
Case Control Study Design
 Data collection : Uniformed questionnaire & direct contact .
( Specific Inclusion & exclusion criteria's ).
 Research Place: the Medical Centres in West Bank
 Ethical consideration: approved by the Research and Ethical
Committee in An-Najah University and Alquds University.
 Sample processing:
1-Blood sample collection ( Three medical centers )
2-Genomic DNA Purification (Master Pure TM Genomic DNA
Purification Kit) .
3-Genomic DNA Qualification (Agarose gel electrophoresis )
10
Genomic DNA Qualification (Agarose gel
.
electrophoresis
1
2
3
4
5
6
7
 Lanes 1-6 contain
separate DNA samples
(1 ul)
 Lane 7 contains 100
Base Pair DNA Ladder
as size marker.
** 
Figure 2.1 Agarose gel electrophoresis
For Genomic DNA
11
Identification of Mutation PCR & ARMS
Test
ARMS Test :One of the basic
methods of DNA assays
It is an accurate, rapid and simple
method (amplification +diagnostic
steps are combines). . ***
The use of allele-specific primers
for PCR amplification
RES
12
Gel electrophoresis for Leiden mutation
PCR product analysis
1
2
3
4
5
6
7
8
Ula Abu Hilal.
9
10 11 12 13 14 15 16 17 18 19 20
13
Inclusion Criteria
Case group
Control group
(191)
(205)
R.M
1ST -
>3
2nd-
>2
IUFD-
>1
PIH
4/13/2015
At least two
pregnancies
No history of any
poor pregnancy
outcomes
14
Exclusion Criteria
Uterine
abnormalities
A known cause
of abortion
Pre-existing
HTN, DM,
Ren. diseases.
Multiple
pregnancy and
IVF
Pre-existing
endocrine
diseases
A known
Leiden
mutation
4/13/2015
15
Results
Obstetric History
Case group: 191 women (at least 3 RA ).
Control group: 205 women (gave normal birth for at
least two babies).
Data indicates that both groups show basically similar
distribution according to Age , BMI values & usage of
OCPs.
The prevalence of Thromboembolic events among
.relatives of the case group (18.5 %) was higher than
that among relatives of the control group (2 %) ( P=
0.006(
16
Characteristics of study participants.
Cases
Age (y)
Marriage age (y)
Regional
Camp
City
Village
Smokers
Education level
Low
High
BMI*
Normal
Overweight
Obese
Abortion
Controls
No
%
31.9
18.8
-
No
32.0
19.3
%
-
115.0
47.0
29.0
19.0
60.0
24.7
15.3
10.0
141.0
40.0
24.0
19.0
69.0
19.4
11.6
10.0
153.0
38.0
79.5
20.5
166.0
39.0
81.3
18.7
55.0
76.0
57.0
78.0
29.2
40.1
30.7
43.0
61.0
63.0
70.0
0.0
30.0
30.7
34.2
0.0
Leiden Mutation Prevalence among participants.
Genotypes
The relationship between Factor V Leiden
mutation and first trimester miscarriages
Mutation
1st trimester
aborters
(n=66)
Controls
(n=155)
Odd Ratio
P-value
Leiden
mutation
14 (21.2%)
13 (8.4%)
2.941
.012
The relationship between Factor V Leiden
mutation and second trimester miscarriages
Mutation
2d
trimester
aborters
(n=25)
Controls
(n=155)
Odd Ratio
P-value
Leiden
mutation
12 (48%)
13 (8.4%)
10.083
.000
The relationship between Factor V Leiden
mutation and IUFD
Mutation
IUFD
(n=26)
Controls
(n=155)
Odd Ratio
P-value
Leiden
mutation
8 (30.8 %)
13 (8.4%)
4.855
.001
The relationship between Factor V
Leiden mutation and PIH
Mutation
PIH
(n=20)
Controls
(n=155)
Odd Ratio
P-value
Leiden
mutation
1 (5%)
13 (8.4%)
.575
1
Conclusion