Professional Documents
Culture Documents
HIV
Hi
chng suy gim min dch ngi Ngun gc 1900 ,1 th sn nhim AIDS virus t Pan troglodytes chimpanzee 1981-2006 ,25 tr ngi 0,4% nhim HIV ,2009
HIV
H
Ritrovirus Chi Lentivirus Di truyn ngc chiu t RNA sang DNA HIV-1 , HIV-2
TI LNG VIRUT
o
s lng bn sao ca virut trong mu Copies/ml Gi tr nn Chu k ngng (CT) ng nn (base line)
CC K THUT PHN T
K thut real time PCR K thut bDNA K thut NASBA
O TI LNG VIRUT
l huyt tng thu RNA tinh sch RNA virut phin m ngc thnh cDNA Mu d Taq man bt cp vi cDNA Taq polymerase ct b mu d , nh sng hunh quang pht ra
KIT TH HIV
1.Khuch i chui acid nucleic ca hn 109 bn cth c thc hin ch trong 90 pht hnh ng ba enzyme. 2. Thit b t tin thermocycling l khng cn thit nh l cc phn ng xy ra isothermally 41 C 3. N gip trong phn ng RT-PCR tt hn v n cung cp ng hc khuch i nhanh hn. 4. Retrovirus c c pht hin v nh lng bng cch khuch i ch c b gen RNA v khng phi bn sao DNA proviral. 5. N c th o sao chp ca virus DNA bng cch pht hin cc biu hin mRNA cui. 6. N h tr vic pht hin cc trnh t mRNA con ngi m khngc cc nguy c nhim DNA
-Vect liu php thit k t adenovirus l mt vect lai cha ba loi gen: gen ca adenovirus l (on CCD Ad5; gen ca HIV l gen gag v tpa bin i ca HIV ; gen ca ngi l promoter CMV (hCMV - gen human cytomegavirus). to nn hai vect liu php gen phng chng HIV l vect Ad5FLgag (Ad5 HIVI gag) v Ad5tpagag (Ad CMVI- ipaHIVgag). Cc vect liu php gen Ad5FLgag vi Ad5tpagag d c th nghim trn kh v trn chui cho kt qu rt kh quan, t l t bo nhim HIV ca kh dc iu tr gim nhiu so vi kh khng c iu tr. Cc vect ny l nhng vect trin vng c th s dng phng v chng HIV/AIDS cho ngi trong thi gian sp ti.
-Vect liu php thit k t virus HIV-1 c thit k v th nghim t nm 1991 (Poznansky v cng s, 1991; Shimada v cng s 1991). C s thit k vect liu php phng chng HIV -1 t b gen HIV - 1 bin di, v c ci gn thm mi s gen khc c chc nng c ch ti bn hoc hot ng ca HIV. n nay, c nhi loi vecto phng chng HIV c s dng. Trong c mt s vect th nghim c hiu qu.
-V d: Vect pCMVAR9 bao gm b gen virus HIV-1 d ct bt mt s gen, v ci thm promoter hCMV ca b gen ngi (hCMV gen human cytomegavirus) v tn hiu ng gi SD (packaging signal); Vect m hoa protein v ca virus (env-coding plasmid), plasmid ny dc thit k gm cc gen m ha v ca virus MLV (Moloney murine leukemia virus); Vecto HIV chuyn dch (transfer vector), vect ny mang cc gen GA, RRE v promoter CMV ca b gen ngi c chc nng ngn cn ti bn gen ca HIV trong t bo ng thi hn ch kh nng gn b gen HIV vi b gen t bo....
-Cc vect HIV-1 phng chng nhim HIV/AIDS c th nghim trn chut, c hiu qu hn ch pht trin ca virus HIV trong t bo. Tuy nhin khi s dng viral vect HIV-1 c hin tng ti t hp gen gia vect liu php vi virus HIV, dn n cc t bin gen v cc hu qu khng mong mun. Do vy vect liu php 111V-1 cho n nay cn l vn cn tip tc nghin cu, th nghim trn ng vt v trn cc t bo ngi in vitro.
hoc c ch hot ng phin m ca nhiu gen khc nhau ca HIV nh c ch gen gag (Lori v cng s, 1994 ), km hm gen rev (Vandedriessche v cng s, 1995) gen em ((uotoku. J v cng s, 1991)... -Ribozvm c dng tm n RNA thng tin (mRNA) ca HIV , thc hin phn ng ct c hiu, lm mt hot tnh mRNA ca HIV (Dropulic, 1994). Virus HIV-1 ribozym c th tc ng v gy mt hot tnh mRNA ca mt s gen HIV nh cc gen gag, pol, tat (Chen v cng s, 1992; Dropulic, 1994)...
vecto phng chng HIV/AIDS, tuy nghin cn th nghim trn ngi thu c kt qu cn thp. Cn nghin cu tm ra cc vect liu c hiu qu thc s, gp phn hn ch tc ng ca i dch AIDS ton cu. -S dng cc liu php gen phng chng HIV/A1DS, cn la chn cc gii php thch hp vi nhng giai on khc nhau trong chu trnh xm nhim t bo v pht trin ca HIV, nhm m bo hiu qu ca liu php.
Tip cn t bo ti th th CD4
Oligo-antisens chng lin kt th th, khng th chng CCR,, ribozym Ribozym, oligo-antisens chng env transdominant gp41
2. Gn vi th th CD4
3. Phn li tch khi phn v 4. Phin m ngc to cDNA 5. B gen HIV gn vi b gen t ho 6. Phin m to mRNA virus 7.Dch m tng hp protein v 8.Hnh thnh v capsid 9. Ti bn b gen H1V 10. Lp ghp cc ht virion 11. Cc virus HIV mi ph t bo 12. Tip tc xm nhim t bo khc
S dng gen t bin Rev 10 - To mt loi pr t bin trong t bo bng gen t bin Rev 10 - S dng vect oligonucleotid mang Rev 10, chuyn vo t bo lympho TCD4 bo nh k thut bn gen. Kt qu cho thy, c s ti t hp vi b gen t bo to nn cc t bo lympho TCD4 mang gen t bin Rev 10, cc t bo ny c kh nng km hm s xm nhim ca HIV (Woffendin v cng s, 1996; Lim v cng s, 1993).
a.
CC GEN CYTOKIN
-Cytokin : l cc pr , cc cht to nn bi h thng t bo min dch trong p ng min dch. -Trong trng hp t bo nhim HIV, do hot dng cc gen HIV lm suy yu kh nng sn sinh interferon ca t bo. Liu php gen vi chin lc a mt gen liu php tng cng hot ng gen sn sinh interferon, v ang c nghin cu v th nghim trn chut. -Gen IL-2 c tc dng kch thch s tng sinh ca cc t bo T CD4 ngn cn s ph hy h min dch do HIV gy ra, khng ch ti bn v nhn ln ca HIV. Vi vai tr quan trng ca gen IL-2 di vi h thng min dch chng HIV/AIDS, hin nay mt s th nghim liu php gen theo xu hng tng cng gen m ha IL-2 cho cc t bo nhim HIV, thu c nhiu kt qu bc u.
-Khi HIV mi xm nhim t bo, bng liu php a cc gen t t (Ced) thymidin kinase (TK) phi hp s dng thuc chng virus ganciclovir (GCV), to thnh phc hp ganciclovir-phosphat c ch hot ng ca enzym DNA polymerase qu trnh ti bn gen ca t bo b ngng ,s phn chia t bo b gin on t bo co li v cht.
S VAI TR CA VACXIN DNA TRONG LIU PHP GEN PHNG CHNG HIV/AIDS
-Hin nay vaccin DNA HIV-1 v ang c th nghim phng chng HIV/AIDS trn chui c hiu qu cao. Bn cht ca vaccin DNA HIV-1 l mt vect liu php gen, c ci tin t b gen HIV-I. Mt s loi vaccin DNA, c th nghim phng chng HIV/AIDS c to thnh t cc plasmid ti t hp, gm plasmid pTXGE v cc gen c hiu cho h thng min dch ngi nh gen IL-2, gen IFN-y v gen GM- CSF. Plasmid ti l hp pTXGE c to thnh t plasmid pTX (4,5 -5 kb) ci thm b gen H1V-1 bin i ( xo b mt s vng gen ca HIV nh gen pol, cc gen vpu, vpr, tat b xa mt vi vng nh).
-Th nghim th nghim vaccin DNA phng chng HIV/AIDS trn chut thnh cng, m ra mt trin vng mi trong iu tr HIV/AIDS cho ngi bng vaccin DNA.
CM N S QUAN TM THEO
DI CA THY V CC BN