: *
! " #
$
%"
$
& ' ( .
*+
:,$ " - ' [email protected]
! " # $ %&
' ( ") *+
Multiplex Nested-PCR
1 2
3 4
*
1 -
! "
#$ % & ' ( )*
+,-.
,- . + + .
2 - 1 2$
,$
( # 3 4
% & ' (,#*
5 6 .7 ( )*
+ ,8
+ + .
3 - * 2$ !) ( 2$ : ;
#$ % (,#*
#$ % & ' ( )*
7 & < + 4 +
.
4 - (
# 3 4
% & ' (,#*
5 6 .7 ( )*
+ ,8
+ + .
! / #
$%&
! / ' 4
/ )
$ 97
,$ $ / 78
: 1 / 8 / 97 :
12 / 9 / 97
:" #
$
% & "' ( )
* '+
, -. "* " /0 1
2 3
% &* 4&
5 "6 7 62 8 8
62
9 : . * ) 0 * < * "=> : ,
8 >? @ 7 "5
$ 3+ "* A 3 ? 61> B7 A C%0 *
>
Multiplex
Nested-PCR , %0 D -(
0 , %2 E F , %2G '* 7 8
.
B7 7 :
"
H 0 E7 1396
=K 400 M0 K A 3 ? "5 A +%2
60 - A 65 N6G 16 E 6'.
, 6
%07 , %0 &
N 7? O ( &
8 P . B7 A C%0 * "5 P
%0 6'Q ) 6 )
62 7 6 ( 7
( Q ) TU
= . -V 7
%' ) / E F ( W X%60 6 .
60 * DNA
> 61>
B7 6*
Multiplex Nested-PCR
8 E F .
"%
:
310 7? O ( "
YX* A
$ 7 ,
, %0 &
N
=K &
6 "
) 5 / 1 (%
A >?
* 60 "
E
7 7 90 ( "
7? O YX* "* ] * A
^ , %0 & /-* 7 W - _
%T` "
+ B 8 6
. 6 ) * 60
8 >?
0 "*
E 2>
E7 a 7 1 0 ) E
7 0 7 E
2>
. 5 A 5 (E7
"F % 7 bc*
8 :
% d , 5 , 1 "
,? "
0 " 8 7 H #%
E 7 0 7 E
62>
E7 6
e X5K
f 3+
$ 07 "*
"' K PCR N 2* 3 1 C
* K X5K f 107 1
. *
Ag 7 ' :
$ 0 , %0 , %2 , %2G '* 7 Multiplex Nested-PCR
"5
? 3 .
" #
A BB8 - BB* BB BB CBB BB D , ,B$ E ; - F . 4 5 " 5@
. B$ , G . ? . HI* (,2*
JB K$ . # ( < L " - MN B2 B L "$ , + . - 5 O P
BB ) "BB BB 5BB 1 2
1 BB BB .(
B L "$ , + . . (,$ * 2 S ? <7 A 2016 A B 2 V' I - 5 L 6
D (. W 216
D 5 6 + ' . B
91
,8 5 " (,$ S * 5
B* + '
A BB . 2015 BB BB Z BB + BB .,BB$ BB 6
,8 445000 A B ? ( + L
- 2015 2016 B D B Z 6
MN B2 B4 5 [ K- 7 \ $ ] 2L MN 2 K B[ & 7 1 B ; ; B$ 5B , (. 8 [7
) " 5 [ 2
) . .(
B B^ "B' 5B D J
(,B$ B B _6B `B # - <
aPB B L Bb ; cB% + BWL - d ) 7 B B 5B + B L 3
BB- + B . B# + B .(
"B ; B$ 5B , 5B P2 D (,$ 5 < 2$
+ BB 5BB 5BB e BB +7 BB D C ,BB MN BB2 B- + $) + + - + f ' +
) ,B$ B (g 2 "[ L 4 5
5B B _,B- .(
M[ D _c8 1404
B' A B hBP; +
B D + , 5 P2 C 5 +,$ ! ,W ? ) ,$
6 7 .(
B L A B 2 5B J - j . # . B# + 2 5 5 " ![ 7 - 5*% A 2 D K- B B2 5B* B - S ? kl ? B; B B + B L V' I e D A 5B B47 !B[ 7 B- 5B > B2 . - 7 . ) B- H< B$ . B# B- +7 ; ,m " . n
4 , % J B- S 5 5 " C 3
B' o B$ B FB 2 B D )B* % 4 2 ) " d 8 9
.(
4 BBB#4 BBB$ VBBB' I BBB- S ) C 3 4 2 10-12
q,2 ( K2 J 7 . [ #4
) . '% ( ^ . B A B ' B 7 (PCR
& BB 6% H IBB*
BB; ( BB BB BB- 5BB*%
) , 13 14 A ,B & B - S . # .(
B* B D !) 5 ![ 7 - 5*% 47 ? @ a
. ( B B (,B$ , Br ( B B- !B[ 7 B; ,m S .,B$ B s # # s # #
B "B; " 8. % c " B 8 . ( B
) "B B< 2 % 13 15
B- S .(
PCR
B ( B Br b < t6 B D B " 8 H I*
10 - 1 - !) 5B ,B + B< 4 #
B- S B % #B # H IB* ,B; u2%
( 6B S ?B ." (,$ 7 v J Id I*
& B - " H I* !) 5 H S . MN B2 D H I* " LPCR
. ( ( F 2 ? % A S ?
B; K B% B B B "B + B ? 2w -
5 . ,@ + - P +
K .7
) ;
13 .(
]7 , B x WB o B$ ? $ ! 4 5 + ; B$ ] B2L ] B2L MN B2 [ yL - ? B2w - + B % + B y[ - B* ^ C + 2 5 D - * ? . A 2 + , L + B .,B$ B "B - z 8 $ , !#*
+ B % * D N . + f ' + ? * ( ) B* D
16 ?B .(
5B*% B47 > B + B 'r _,- 5@4 P
@ - 5*% & 6% 5 > $ ? ... !;
, 8 + # -
. ( B B B D !) " . n 5 ( ![ 7 - 4 BB#4 S
Nested-PCR Multiplex
+ BB $
$ C ." (,$ & ^ + f ' + +
B7 7
MN B2 @P r r 5 d 8 5@4 P B$ CB + B $ B D B A P< %
A B & 5B + B f ' + B + 1396
W + $ - 7 ., & ^
"BB2 . ,BB . 5BB BB + BB- BB$D g BB2[
- FI 2 - 15
!B @ B 5B C .
B .,B , , r ( < + < ![ 7 - 5*% (, 7 ) B , Br B- S . ![ 7 - 5*% , r
Hand
catch
B- 54 4 . ( 8 ) '< ? ( n 7 (,2#
2Bb B@ WN L < ?
)
Pit-shelter
B (,$ , r - 5 .,$ ( (
& [ ' 5 5 *m7 Cf 5W2% C . ( n 7 5 B* "B[ B B; # B 6% - + 4 5 5B 7 hB L B ' & B . ,@ 5'r [6 ., ,$
8 - 54 4 !< (,$ , r - !#4
70 ; ,r
B .,B2 [ A B 5BN "B$ , 5 (,$ ( !B[ 7 D B B Z 2$ ( *8 s 4 . (
B- ] B !B< 5B (,B$ ,L ( < + < - 5
/ 1
.,$ g ] 5 bI* A 4 ' ( | b < [ 8 5 , C 5 - 5 5B .,B$
B ,B B47 5B - . 5 ] !< - g I + . [ DNA
°C
20 - ; ,2 [ .
N -1K PCR
g I DNA
g I " . ( . DNA
"BB . % # " BB$ ,BB 4 BB S "BB[ + BB<
)
DynaBio Blood/Tissue Genomic DNA Extraction
^ (Kit
C 2# ."[ &
Multiplex Nested PCR
B B- % . ( B B & B 6% B- 5 ? @
A ,L) (,$ 8 N 1
"B L 5B@4 P ?B .,$ & ^ (
K2BB -PCR
1 ,BB }' ) 4 BB % "BB L CBB .
6rPLU
5rPLU
K2B .,B$ ( -PCR
2 .
BB- %
rFAL.1 rFAL.2
rVIV.1
.rVIV
2 "BB L
.,B$ ( B & B Br 5B & B 6% 5 ? @ A b PCR
"W 5 P ]7 o A 1
5B 100
)4 + 2 5 (,$ M ; ., ( & PCR
!8 A #4 . ,L :,$ . O $ 5 PCR
8 DNA
95 , 5 5L 5
5 ; B8 94
, 5 5L 30
) 5 k 25 A 5'8 !#
30
!#
Nested
,B ,L B 4 #4 "< B > B$ (
58 DNA
, 5 5L 1
5 ; 72 ,B 5 5L
60 A 5'8 5 k 30
B 5 k
Nested
B &
. n S 72
, 5 5L 10
5 ;
A 5BB'8 5
BB 5BB ;
Nested
& BB "BB .
5 K- K2 20
) 5L 14 17 & B . •B% .(
K2 . BG B 5B@P; B €# B . " L PCR
. 7 A3 . [ #4 S 5
/ 1
,B ( B %
) 18 19 .(
"%
> BB ^ 400
4 BB 5BB ( BB !BB[ 7 5BB*%
, , r •
. . 310 . (,B$ 7 h L 5
B$ CB + B $ + - $D - KI ,@
6 ) 5 5 / 1 • B & 6% 5 ( 47 (%
.
90 g B2[ B- KI 5 e (,$ 7 h L 5
., ) S W€ 5 ‚ - c + $ "2 B 6% 5B 47 ? & n B 4 [ &
(& % 4 [ & 6% • & 6%) •#
> B ^ .,B* (,- * 66
/ 1 4 B B- !B[ 7 . %
., & 6% !) 5 ( 47 (,$ •
H7 ( 1 .
* Multiplex Nested- PCR E 0 " 8 ) =K /&(
• [ A b A N
4
% &
14 1200bp
5'-CTTGTTGTTGCCTTAAACTT-3'
5'-TTAAAATTGCAGTTAAAACG-3' rplu5
rplu6
14 205bp
5'-TTAAACTGGTTTGGGAAAACCAAATATATT-3' 5'-ACACAATGAACTCAATCATGACTACCCGTC-3'
rFAL.1 rFAL.2
14 120bp
5'-CGCTTCTAGCTTAATCCACATAACTGATAC-3'
5'-ACTTCCAAGCCGAAGCAAAGAAAGTCCTTA-3' rVIV.1
rVIV.2
H7 ( 2 .
& N , %0 & 3 ? "5 E 0 "* 8 >? Y- d %
! ,@ Nested-PCR S 5 (,$ 2$ & 6% 5 $ C + $ *I
2 & n 4 [ & 6% • & 6%
30 30 0 0 ,@ "2
100% 100% 0 0 ,r
42 42 0 0 ,@ g 2[
100% 100% 0 0 ,r
90 89 0 1 ,@ $D
100% 98/9% 0 1/1 ,r
96 94 0 2 ,@ + -
100% 97/02 0 2/08 ,r
124 121 0 3 ,@
100% 97/58 0 2/42 ,r
31 1 . H Vc Y 8? Hg 7 *PCR
E 0 3+ h *
.
) $ 1 7 2 7 3 ) $ /T` H %- 10
- 5
7 E 0
120bp) ) $ ( 4 100bp
7 bc*
"F % 8
BB D BB BB + BB2 5BB . BB2-
? !#BB*
$ , 8 5
; B$ ] B2L B
+ 5B B (j + B
+ + B f ' O BP
( B
B KB
BB$
o BBb ; BB L
BB
? & BB 5BB P2 BB
,
@ " L d 8 5@4 P .
?
$
>
I*
H BB 6% 5BB
&
BB- 5BB*%
7
!BB[
S . !BB;
4 #4
Nested-PCR Multiplex
( B B
, . B
?
S BB 8
"
BB D BB BB ( 6BB ( BB
I*
H !) 5 •2L I*
H - "
•#
) 20 .(
u 2 5@4 P 5 + *
5 + $ ![ 7 5*% F4 m B
C + B B$
@ - 5*% & 6% 5 > $ ? ... !;
, 8 + # -
+ 4 ![ 7 + f ' B
• ) 33 / 83
% (
,$
) A ,L 2 (.
D 5 F4 m
+ B
•
"
) 21 7 ( + ( 4 5
/ 1 + $ - 5*% % C
$
? 5'}
, 5 ,2 5B [ ?B B B
u
. !r 8 @4 P
"B P .
5B@4 P
; * o 5 5 * B# -
+ B- + B $
B f BB*
& BB^
B , 5BB FBB4 m BB
?
BB $ BB
BB
• BB ) 22 BB2w - .(
? BB 7
+ # - 5@4 P MN 2
, C + ( B B
S . ., & ^ PCR
u
? B8 5B@4 P +7 .
6% 5 &
• B 5B P2 FB4 m !B)
,$
) 23 .(
+ # - 5@4 P
Bf . B
?
S . (
Nested-PCR
5B ,B & ^
BB u BB 6% + BB*
&
BB
• FBB4 m !BB)
5 P2 5@4 P
,$
) 24 .(
Br 5B@4 P
oBB 5B [ vBB f C BB BB
( BB + BB# -
2L 7 / 5 % - 5*%
![ 7 ' 2 S ( 47PCR
6% 5
&
• ) , 25 (.
@4 P 5 6 + # - B $ 5B
2 "B[ Br 1 B[ + J 512
5B*%
S 5BB ( BB !BB[ 7 PCR
@ BB
? , ,BB$ 5BB .
BB u
5@4 P 5 + * !B[ 7 (,$ ƒ MN 2 F4 m
4 ) 38 / 71
% ( - (
‚ 47 6% !) 5
&
S PCR
% 5*
- ) S , 4 8 . 5B
d 8 5@4 P 66
/ 1
% - 5*% . 5B 5B@4 P ![ 7
6%
&
+ ., ( 47 +
„ L + f '
MN BB2 ,BB C D BB BB ] BB BB BB$
, 5
? 5B (,B$ `B 1 B[ + B B + B
47 6% 5
&
- ![ 7
? 2 S B 5 P
$ ) 26 .(
5@4 P ,2- 5 *
% - " L + # -
47 5*%
- B 6% 5 ![ 7
&
523 5B*%
, ; 5@4 P ![ 7 .
u
? + B* 5B@4 P
5 5 D 7 ![ 7 F4 m
• . •B% 5 ,$
. ,L B; ,m
B
#
% #B 7 B- B
‚
. n ) (,- *
, 4 8 . 5#
S B
PCR
7 D B 7 !B[ 7 5*%
• ) 4 / 3 B 6% 5B ( B47 (%
&
4 [ , & % )
27 .(
u 5@4 P ) oB 5B
?w n + # - + B* ,B$ & B^ #
) 5 / 9
![ 7 (%
- 7
( 4 6% 5
&
•
6%
&
4 [ B & % ,2B$
) 28 .(
•4 B
5@4 P + # - W ' A $
S . ( B B
Nested-PCR
5B 5B , & ^
!B[ 7 FB4 m B-
W47 1 83
% - 5 ( 47 6% >
&
4 [
& %
• , ) 29 .(
$7 + B# -
5@4 P S . ( ,2- ] m
Nested-PCR
47
![ 7 - ,
u + B* + 7
5 8 / 2 - ![ 7 %
% ] 1 # B 6% 5 ( 47
&
4 [ & %
• 47 , B Br 5
•#
(,- * ,
) 30 .(
u f B$ G . d 8 5@4 P \ [ 5@4 P
>
- 5 6%
&
$ .,$
>
B- 5
6% V' I
&
kxB "B + L V' I MN 2
! B$ B- ]7 B 5B B;
o
3 4
# ,
? 5'}
.,$ kl
- BBB 7 .
BBB F BBB- 5BBB !BBB[ 7
BBB +
. nBB
"
BB; ,BBm S CBB 5BB BB 7
BB-
5 " L 1 8 ?} P
% )*
B
D 8
cB% ] 2 L B
,B$
) 9 .(
B
?
+ B* 5B@4 P S 5B
Nested PCR
-
Multiplex
"4 5 + ,@
. 5
. (
? S .
* 5 a B; ,m !B[ 7
"B
8 - 5
,; (,$ C*<
B B + B
2 . ;
? .
^ 5 .7 ( )*
& -
,B 7 MN B2 5 !2 % C
B 5B ( BW B-
h L 7 5 P2 (,$
.7 + B ( )B*
B-
.7 K . ;
L BB
? BBy[
BB % L
BB 5BB BB
+ $
C $ + + + f ' "[
4 B8 t ,BW B* 5B + $, 7 B & B^
B 5B
B P< % + B2 5B + B % + y[
? - B*
D A , (,$ 5 < 2$
) 31 21 2 ( 5B@4 P .
^
# - + y … B2 + $ B
B
D [ K A
1387 B†8 5
> W 5 ) ,B "WB )
32 z .(
B K B# -
5@4 P
*
? m - $ 5 e
L J-
"4 + B % + f ' + B
+ B
,2 BB + BB f ' )
33 + BB# - BB% .(
BB 5BB
- [
D K- k 1 W ,2 $
5 < % 5
? 5^
, , 5B B
? B$
4 5B B
!
c% L > BW ,B 7 "B[ + B 5B )
( B -
% BP< E @ B
, B- D B B ,B$
) 34 .(
cB4
- 5 +
"
B2 5B L +
B*4 f + B
D A 2 J . ( $
% @4 P ,
3 4 C D 5B " ( + * B
+ D _6< B B
[7 B - B*
B) h B
$ o ';
( B C 5B 5B*
B2 ,B$ B
) 35-37 , .(
? 2@
5
?
@d
"
% , " ?#
^ ) [ 5 ,$ k K
, MN B2
8 P< % A
$
>
* ,B$ ( -
) 38 .(
+ + B$ B ( 6 + f ' o
';
B
L ! Bb ;
B)2- [ B
"
B
"
L A 5 A B- 5
D A B 2 B
@;
B
"
. - ) - B* B + B % + B y[
y[ + L 5 e `8 W
% . B
4 5
? K
, ) 31 .(
5B 5BL
5B#2 + B + B
+ B f '
D _c8 5'8 ;
# .
? B k
?
S BB% "BB L BB- K
BB% \ BB[ 5BB K
BB47
5*%
- ( ![ 7 B
B .,B$
u 5B@4 P . !Br 8
d 8 5
'W; @4 P B47 5B + B*
BB- 5BB*%
D !BB;
BB + BB $ BB
C BB$
BB
@r G 5 5 $
, 5 _cB8 D B
5 P2 A
1404 .
@4 P 5
B ,
* .
? u + * 5 ,-
- + # +7 5B
5 - A
• 4 [ B ‡ & B%
,B-
I*
H ') D S , PCR
C ! #
, IB*
H
% #B # .,B$
B2
?
% @4 P + ,
3 4 C 4 B#4 BN A B B-
& ^ 5 5L . 5#2
+ + + f '
D _c8 5'8 B
B) ! B 5B - B;
. 52 A 2
?
% 5 , ,
? B
. $ ^2 B4 2 ,; " &.D
& ,B BN 5B
( W & (,$ A * MN 2
* B . (,B$ A 2 B
5 L B
?
'L 7 ! @
, .
j 7 15K
BB
? BBL !BBr 8 5BB@4 P BB*I
% . BB + 5BB
2$
J < ,$
• ,B ( B
?
5' . B# -
% & B' ( )B* 5 ,2 IB
#B$
+ ,BB- . !2BB % BB +7
8 BB
"
BB ^4 C BB<
h L 7 5B
IB*
H D B 5B*%
B- !B;
,;
.
- 5*% & 6% 5 > $ ? @ ...!;
, 8 + # -
References
1. D’Acremont V, Bosman A. WHO informal consultation on fever management in peripheral health care settings: a global review of evidence and practice. Geneva:
World Health Organization. 2013:759-764.
2. Organization WH. World malaria report 2016. Geneva: World Health Organization;
2016. Licence: CC BY-NC-SA 3.0 IGO [cited 2017 Jun 13].
3. Reiter P. Global warming and malaria:
knowing the horse before hitching the cart.
Malaria Journal. 2008;7(1):S3.
4. Beljaev A. The malaria situation in the WHO eastern Mediterranean region.
Meditsinskaia parazitologiia i parazitarnye bolezni. 2000(2):12-15.
5. Soleimani-Ahmadi M, Vatandoost H, Zare M, Turki H, Alizadeh A. Topographical distribution of anopheline mosquitoes in an area under elimination programme in the south of Iran. Malaria journal.
2015;14(1):262 (In Persian).
6. Soleimani-Ahmadi M, Vatandoost H, Shaeghi M, Raeisi A, Abedi F, Eshraghian M, et al. Field evaluation of permethrin long-lasting insecticide treated nets (Olyset®) for malaria control in an endemic area, southeast of Iran. Acta tropica.
2012;123(3):146-153(In Persian).
7. Elimination M. In Islamic Republic of Iran (Horizon 1404). Ministry of Health and Medical Education.
8. Raghavendra K, Barik TK, Reddy BN, Sharma P, Dash AP. Malaria vector control:
from past to future. Parasitology research.
2011;108(4):757-779.
9. Soltani A, Vatandoost H, Jabbari H, Mesdaghinia A, Mahvi A, Younesian M, et al. Field efficacy of expanded polystyrene and shredded waste polystyrene beads for mosquito control in artificial pools and field trials, Islamic Republic of Iran. 2012 (In Persian).
10. Mirahmadi H, Fallahi Sh, Taghipour N, Turki H, Seyyed Tabaei SJ. Cloning and Sequence Analysis of Recombinant Plasmodium vivax Merozoite Surface Protein 1 (PvMSP-142 kDa) In pTZ57R/T Vector. Iranian journal of parasitology.
2015;10(2):197-205 (In Persian).
11. Mirahmadi H, Fallahi Sh, Omrani VF, Kazemi B, Haghighi A, Tabaei SJ. High- Level Expression of Immunogenic Recombinant Plasmodium vivax Merozoite Surface Protein (Pvmsp-142 kDa) in pGEX 6P1 Vector. Iranian journal of public health.
2015;44(1):89 (In Persian).
12. Mirahmadi H, Fallahi Sh, Seyyed Tabaei SJ. Soluble recombinant merozoite surface antigen-142kDa of Plasmodiumvivax:
Animproveddiagnostic antigen for vivax malaria. Journal of microbiological methods. 2016;123:44-50 (In Persian).
13. Mohammadzadeh T, Hatam G, Kalantari M, Sarkari B, Motazedian MH, Sadjjadi SM, et al. Molecular and microscopic-based characterization of Plasmodium spp. in fars and Hormozgan Provinces, South of Iran.
Journal of tropical medicine. 2014;2014 (In Persian).
14. Pinheirob VE, Thaithongc S, Browna KN.
High sensitivity of detection of human
malaria parasites by the use of nested polymerase chain reaction. Molecular and biochemical parasitology. 1993;61:315-320.
15. Arez A, Lopes D, Pinto J, Franco A, Snounou G, Do Rosário V. Plasmodium sp.:
optimal protocols for PCR detection of low parasite numbers from mosquito (Anopheles sp.) samples. Experimental parasitology. 2000;94(4):269-272.
16. Halimi M, Delavari M. Jafarymadrak. M.
Modeling the spatial spread of malaria in Baluchistan province geographically weighted regression model. Journal of spatial planning. 2013;17(3):85-107 (In Persian).
17. Fuehrer H-P, Noedl H. Recent advances in detection of Plasmodium ovale:
implications of separation into the two species Plasmodium ovale wallikeri and Plasmodium ovale curtisi. Journal of clinical microbiology. 2014;52(2):387-391.
18. Zakeri S, Najafabadi ST, Zare A, Djadid ND. Detection of malaria parasites by nested PCR in south-eastern, Iran: evidence of highly mixed infections in Chahbahar district. Malaria journal. 2002;1(1):2 (In Persian).
19. Brown AE, Kain KC, Pipithkul J, Webster HK. Demonstration by the polymerase chain reaction of mixed Plasmodium falciparum and P. vivax infections undetected by conventional microscopy.
Transactions of the Royal society of tropical medicine and hygiene. 1992;86(6):609-612.
20. Nankabirwa JI, Yeka A, Arinaitwe E, Kigozi R, Drakeley C, Kamya MR, et al.
Estimating malaria parasite prevalence from
community surveys in Uganda: a comparison of microscopy, rapid diagnostic tests and polymerase chain reaction.
Malaria journal. 2015;14(1):528.
21. Manouchehri A, Zaim M, Emadi A. A review of malaria in Iran, 1975-90. Journal of the American mosquito control association. 1992;8(4):381-385 (In Persian).
22. Oshaghi M, Moradi M, Taghilou B. specific detection of MALARIA parasites using nested-PCR in individual mosquitoes and infected bloods in CHabahar and Iranshahr, Iran. 2004 (In Persian).
23. Assmar M, Ter Hovanessian A, Naddaf S, Piazak N, Masomi H. PCR Detection of malaria parasites in Anopheles stephensi and Anopheles culicifacies mosquitoes collected from southern endemic foci of Iran. Journal of school of public health and institute of public health research.
2005;3(3):19-26.
24. Yu G, Yan G, Zhang N, Zhong D, Wang Y, He Z, et al. The Anopheles community and the role of Anopheles minimus on malaria transmission on the China-Myanmar border.
Parasites & vectors. 2013;6(1):264.
25. Chang KS, Yoo D-H, Ju YR, Lee WG, Roh JY, Kim H-C, et al. Distribution of malaria vectors and incidence of vivax malaria at Korean army installations near the demilitarized zone, Republic of Korea.
Malaria journal. 2016;15(1):259.
26. Kalantari M, Soltani Z, Ebrahimi M, Yousefi M, Amin M, Shafiei A, et al. Monitoring of Plasmodium infection in humans and potential vectors of malaria in a newly emerged focus in southern Iran. Pathogens
- 5*% & 6% 5 > $ ? @ ...!;
, 8 + # -
and global health. 2017;111(1):49-55 (In Persian).
27. Mahapatra N, Marai N, Ranjit M, Parida S, Hansdah D, Hazra R, et al. Detection of Plasmodium falciparum infection in anopheles mosquitoes from Keonjhar district, Orissa, India. Journal of vector borne diseases. 2006;43(4):191.
28. Nepomichene TN, Tata E, Boyer S. Malaria case in Madagascar, probable implication of a new vector, Anopheles coustani. Malaria journal. 2015;14(1):475.
29. González C, Molina AG, León C, Salcedo N, Rondón S, Paz A, et al. Entomological characterization of malaria in northern Colombia through vector and parasite species identification, and analyses of spatial distribution and infection rates.
Malaria journal. 2017;16(1):431.
30. Kumar A, Hosmani R, Jadhav S, Sousa T, Mohanty A, Naik M, et al. Anopheles subpictus carry human malaria parasites in an urban area of Western India and may facilitate perennial malaria transmission.
Malaria journal. 2016;15(1):124.
31. Sartipi M, Khosravi A, Khalaji K, Shamsipour M, Kazemi GMH, Sakeni M, et al. Examining the risk factors of MALARIA:
a matched case –control study. 2014 (In Persian).
32. Raiesi A, Nejati J, Ansari-moghaddam A, Sakeni M, Faraji L, Paktinat B, et al. Effects of foreign immigrants on malaria situation in cleared up and potential foci in one of the highest malaria burden district of southern Iran. Malaria journal. 2012;11(1):P81 (In Persian).
33. Raeisi A, Nikpoor F, Ranjbar Kahkha M, Faraji L. The trend of Malaria in IR Iran from 2002 to 2007. Hakim research journal.
2009;12(1):35-41 (In Persian).
34. Poudat A, Ladoni H, Raeisi A. Possible factors affecting the malaria situation in the city of Bandar Abbas in 1999-2002. Journal of Hormozgan University of Medical Sciences. 2006;2:101-110. (In Persian) 35. Mirahmadi H, Shahrakipour A, Mehravaran
A, Khorashad AS, Rahmati-Balaghaleh M, Zarean M. Evaluation of malaria multiplex/nested PCR performance at low parasite densities and mixed infection in Iran: A country close to malaria elimination.
Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases.
2018;65:283-287 (In Persian).
36. Rahmati Balaghaleh MZM, Afzal Aghaee M, Shamsian SA, Mirahmadi H, Arya A.
Comparison of PfHRP-2/pLDH RDTs with Light Microscopy in a Low Prevalence Setting in Southeastern Iran, Sistan and Baluchestan: Due to Implementation of Malaria Elimination Program. International Journal of Infection 2018;5(1):e12286 (In Persian).
37. Ebrahimzadeh A NDS, Mirahmadi H, Mehravaran A, Salimi Khorashad A, Turki H. The Incidence of Current Infection with Different Human Malaria Species by Polymerase Chain Reaction for Diagnosis of Suspicious Malaria Patients on Elimination Region Sistan and Baluchistan Province, Southeast of Iran. Jundishapur
Journal of microbiology. 2017;10(10):
e58254 (In Persian).
38. Soleimani Fard SD, Akbari M, Sabet Ghadam M, Saberi S. Malaria situation in Isfahan in the last five years. Journal of Isfahan Medical School, 2012;29(132):
173-280 (In Persian).
- 5*% & 6% 5 > $ ? @ ...!;
, 8 + # -
Determination of the prevalence and type of plasmodium in mosquitoes present in the city of Nik Shahr, Southern Sistan and Baluchestan
Province, IR Iran, by Multiplex Nested-PCR
Mirahmadi M1, Yazdani A2, Fallahi Sh3, Kazemipour N4٭
1.Assistant Professor, Infectious Disease and Tropical Medicine Research Center, Tuberculosis Resistance Institute, Zahedan University of Medical Sciences, Zahedan, Iran.
2.Graduate Student, Department of Microbiology, Faculty of Basic Sciences, Islamic Azad University, Kerman, Kerman, Iran.
3.Associate Professor, Department of Parasitology and Mycology, Faculty of Medicine, Lorestan University of Medical Sciences, Khorramabad, Iran.
4.Assistant Professor, Department of Microbiology, Faculty of Basic Sciences, Islamic Azad University, Kerman, Kerman, Iran, [email protected].
Received: 23 Oct 2018 Accepted: 3 Dec 2018
Abstract
Background: Malaria is one of the most important parasitic diseases, and is considered one of the important health issues in tropical and subtropical countries. The aim of this study was to determine the rate and type of infection of Anopheles mosquitoes by malarial parasites using Multiplex Nested-PCR in the South of Sistan and Baluchestan province.
Materials and Methods: In the second half of 2017, 400 Anopheles mosquitoes were collected from Zahedan Medical Insecticide Check Centers located in villages around Nikshahr city. The mosquitoes were caught by hand-held methods in domestic (human and animal), natural and artificial outdoor places (Shelterpit). After DNA extraction, molecular analysis was performed using Multiplex Nested-PCR.
Results: Of the 310 samples collected from parts of Lashar, Ahuran and the centre of Nik Shahr city, 6 samples (1.5%) were found to be infected with Plasmodium vivacs, and 90 samples collected from the Fennoj and Bennett sections of the city had no infection. Samples containing plasmodium falciparum and a mixture of Plasmodium vivacs and Plasmodium falciparum were not detected in this study.
Conclusion: The results show that in places where the transmission of both species of Plasmodium vivax and Plasmodium falciparum occur, the detection of malarial parasites by PCR can be a very useful complement to confirm microscopic diagnosis.
Keywords: Malaria, Sistan and Baluchestan province, Multiplex Nested-PCR, Anopheles mosquitoes
*Citation: Mirahmadi M, Yazdani A, Fallahi Sh, Kazemipour N. Determination of the prevalence and type of plasmodium in the mosquitoes of the city of Nickshahr in Southern Sistan and Baluchestan province by Multiplex Nested-PCR. Yafte. 2019; 20(4):63-73.
