Validitas Multiplex Real Time Polymerase Chain Reaction

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REAL TIM E PCR

KELOMPOK 9

DAYU PRATAMA SAYUTI

DEWI RATIH

DYAH AYU K 

FIKA YUSTISI AMALIA

NITA MEITHIYASARI

NUR FADILAH

RIA PUSPITA

SUSI SUSANTI

 TRI HANDAYANI

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Pengertian Real Tim e-PCR

Secara U m um

 Real-Time PCR atau jua !ia"a

#i"e!ut $-PCR %$ua&titati'e PCR(me)u*a+a& "e!ua, met#e u&u+me&am*li.+a"i DNA/

 Te+&i+ i&i #iu&a+a& u&tu+me&am*li.+a"i%mem*e)!a&0a+("e+aliu" +ua&ti.+a"i %me&,itu&(

 jumala, ta)et mle+ul DNA ,a"il

am*li.+a"i te)"e!ut

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Real time PCR !e)!e#a #e&a& PCR+&'e&"i&al/ Pe)!e#aa& Real Time PCR

#e&a& PCR +&'e&"i&al 1u+u* "i&i.+a&2#a)i ,al "e&"iti.ta" #a& +ua&tita"/

Pa#a PCR t)a#i"i&al2 1a)a #ete+"ime&u&a+a& ele+t)3)e"i" el !ai+aa)"a mau*u& *lia+)ilami#2 "e#a&+a&*a#a $-PCR #ete+"i me&u&a+a& "0"tem

4u)e"e&"/ Selai& itu *e&u&aa&4u)e"e&" i&i jua mem!e)i+a& "e&"iti.ta"0a& ti&i !ai $-PCR/

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Prinsip Kerja Real Tim e PCR

Real Time me&i+uti *)i&"i* umum)ea+"i PCR2 utama&0a a#ala, DNA0a& tela, #iam*li.+a"i #i,itu&"etela, #ia+umula"i+a& #alam )ea+"i"e1a)a )eal time "e"u#a, "etia*"i+lu" am*li.+a"i "ele"ai

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Alat real tim e PCR

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Skem a pem eriksaan dengan real tim e pcr

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Kurva Real tim e PCR

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VALID ITAS M U LTIPLEX REAL

TIM E PO LYM ERASE CH AIN

REACTIO N 

U N TU K D IAG N O SIS

LIM FAD EN ITIS TU ERKU LO SISPAD A SPESIM EN LO K PARAFIN

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Latar Belakang

Lim3a#e&iti" tu!e)+ul"i" me)u*a+a&tu!e)+ul"i" e+"t)a*a)u 0a& *ali&"e)i& #itemu+a&/ Peme)i+"aa&,i"t*atli ti#a+ mem!e)i+a&i&3)ma"i etili *a"ti *e&0e!a!lim3a#e&iti"2 *a#a,al lim3a#e&iti"

#a*at #i"e!a!+a& Mycobacteriumtuberculosis mau*u& Mycobacteriumnon tuberculosis yang sangat berbedaregimen terapinya.

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K&.)ma"i #ia&"i" te)"a&+a lim3a#e&iti" T5mem*e)u&a+a& *eme)i+"aa& !a+te)i ta,a&a"am %5TA(2 ,i"t*atli2 atau me&u&a+a&te+&i+ real time polymerase chain reaction (RT-PCR).

Saat i&i te+&i+ RT-PCR #i*e)u&a+a& u&tu+me&#ete+"i Mycobacterium/ Peme)i+"aa&multiplex RT-PCR %M RT-PCR( me)u*a+a& alte)&ati3*eme)i+"aa& lim3a#e&iti" T5 "elai& "e1a)a

,i"t*atli2#a& tela, #ia*li+a"i+a& u&tu+#ete+"i "i+ue&" Mycobacterium deoxyribonucleicacid (DN) *a#a !e)!aai !a,a&/

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me&u&a+a& te+&i+ M RT-PCR i&i le!i,a+u)at2*)"e" am*li.+a"i #a& jua

a&ali"i"&0a te)ja#i !e)"amaa& "e,i&a6a+tu 0a& #i!utu,+a& u&tu+i#e&ti.+a"i DNA !a+te)i a+a& le!i,

"i&+at2 #a& #a*at me&#ete+"i 787!a"il #i #alam !e)!aai "am*el +li&i"

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M etode

Se!a&0a+ :; !je+ *e&elitia& 0a& te)#i)i ata"< !l+ *a)a.& lim3a#e&iti" T5 #a& ;; !l+*a)a3i& +elm*+ +&t)l2 #iam!il = > !l+*a)a.& #a)i "etia* ma"i&-m"i& !l+ *a)a.&2

#ila+u+a& e+"t)a+"i DNA #a& *eme)i+"aa&multiplex real time polymerase chain reactionmengguna!an *)ime) e& "i+ue&" "i"i*a&"#$%%& dan gen Mycobacterium protein

tuberculosis MP'$ u&tu+ me&#ete+"i DNA M.tuberculosis dan gen %$# rRN untu!mendete!si Mycobacterium non tuberculosis

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asil

Pa#a *e&elitia& te)#a*at "e!a&0a+< +a"u" #i#ia&"i" "e!aailim3a#e&iti" T5 "e1a)a ,i"t*atli2"etela, #ila+u+a& *eme)i+"aa& M RT-PCR #i#a*at+a& ? +a"u" #i"e!a!+a&le, M. tuberculosis dan %& !asus

negati* bu!an #i"e!a!+a& le, M/tuberculosis maupun MNT.

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Ha"il &eati3 *al"u M RT-PCR *a#a !l+*a)a.& #a*at #i"e!a!+a& le, !e)!aai3a+t) a&ta)a lai& a#ala,

#i"t)i!u"i Mycobacterium yang tida!me)ata *a#a "am*el ja)i&a&

  &e+)"i" 0a& lua" *a#a ja)i&a&Mycobacterium

  proses +!sasi dan *em!uata& !l+

*a)a.&"e)ta *e)u!a,a& "t)u+tu) DNA

"e6a+tu *)"e" .+"a"i 3)mali&/

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F)ame& DNA 0a& #a*at #ia&ali"i"!ia"a&0a mem*u&0ai !* @= !*2 le,

+a)e&a le!i, mu#a, !e)3)ame&ta"i/U+u)a& am*li+& me)u*a+a& 3a+t)0a& mem*u&0ai *e)a& #alam

+e!e),a"ila& am*li.+a"i2 +a)e&a DNA*a#a !l+ *a)a.& "e)i& te)#e)a#a"ile, *)"e#u) .+"a"i #a& embedding

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Ne+)"i" *e)+ijua& 0a& lua" #a*atme&0e!a!+a& ,a"il &eati3 +a)e&a

Mycobacterium ,arang tumbuh di !aia&te&a, ja)i&a& &e+)ti+/ Dae)a, &e+)"i"+e+u)a&a& +"ie& 0a& #i*e)lu+a& u&tu+*e)tum!u,a& !a+te)i2 "e,i&a Mycobacterium

itu le!i, "e)i& !e)a#a #i !aia& te*i &e+)"i"0a& ma"i, #itemu+a& "el ma+)3a/

Ha"il &eati3 *al"u ,i"t*atli *a#a M RT-

PCR #a*at #i"e!a!+a& le, jumla, DNA 0a&+u)a& #a)i 2; > "e,i&a ti#a+ #a*atte)#ete+"i le, alat/

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Pem !a"asan

Lim3a#e&iti" "e)i& #i"e!a!+a& le,!a+te)i M. tuberculosis maupun MNT.

Lim3a#e&iti" T5 *e)lu #i!e#a+a&#e&a& MNT +a)e&a te)a*i&0a 0a&"a&at !e)!e#a "e,i&a #i*e)lu+a&"uatu *eme)i+"aa& 0a& #a*at

me&e&tu+a& etili *e&0e!a!#e&a& *a"ti/

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Multiplex RT-PCR merupa!an salah satualat *eme)i+"aa& 0a& #a*at

me&e&tu+a& etili *a"ti *e&0e!a!lim3a#e&iti"/

I&3e+"i M.tuberculosis merupa!an

 penyebab limadenitis 0a& *ali&"e)i& te)ja#i *a#a a&a+ #a&#e6a"a/Mycobacterium tuberculosismempunyai daya *ate&ita" 0a&

le!i, ti&i #a)i*a#a MNT2"e,i&a#a*at me&im!ul+a& *e&0a+it*a#a*e&#e)ita #e&a& imu&ita" &)mal

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Ha"il &eati3 *al"u M RT-PCR #a*at#i"e!a!+a& le, jumla, +uma& 0a& "e#i+it

%*au"i!a"ile)( #alam ja)i&a&/ Fi+"a"i ja)i&a& #e&a& me&u&a+a&

3)mali& #a& #i"im*a& #alam 6a+tu lama#a*at me&0e!a!+a& *e)u!a,a& "t)u+tu)

#a& #e)a#a"i DNA/ Fi+"a"i 0a& "e!ai+&0a #iu&a+a& u&tu+

,a"il *timal a#ala, *e&u&aa& 1ai)a&neutral buer ormalin #i!a&#i&+a&#e&a& 7 non-buer ormalin +a)e&aa+a& mem*e)lam!at *)"e" #e)a#a"i 0a&

#i"e!a!+a& le, 3)mali&/

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Pe&u&aa& 7 non-buer ormalin

dapat menyebab!an ti#a+te)am*li.+a"i&0a DNA e& "#$%%& padareal time PCR sehingga a!anmemberi!an hasil &eati3 *al"u/

Wa+tu .+"a"i 0a& !ai+ u&tu+ ja)i&a&0a& a+a& #ila+u+a& *eme)i+"aa& MRT-PCR a#ala, ti#a+ le!i, #a)i ;< jam

u&tu+ me&,i&#a)i .+"a"i !e)le!i,a&0a& #a*at me&0e!a!+a& crosslin!dari mole!ul DNA/

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5e!e)a*a 3a+t) 0a& #a*atmeme&a)u,i #e)a#a"i DNA *a#a !l+

*a)a.& te)ma"u+ lama&0a .+"a"i2 *H2+&"e&t)a"i a)am2 #a& "u,u/ O+"i#a"inon-buer 3)mali& mem!e&tu+ a"am

#a& li&+u&a& a"am me&0e!a!+a&#e)a#a"i a"am &u+leat +a)e&a i+ata&B li+"i#i+ *a#a !a"a *u)i&te),i#)li"i" *a#a *H </

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Peme)i+"aa& multiplex real time

 polymerase chain reaction dapatdian,ur!an untu! !on+rmasi #ia&"i"lim3a#e&iti" tu!e)+ul"i" *a#a +a"u"te)"a&+a +uat lim3a#e&iti" T5 "e1a)a

+li&i"2teta*i ,a"il ,i"t*atli ti#a+mem*e)li,at+a& lim3a#e&iti" T5/

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TERIM AKASI