treatment of Juvenile Laryngeal Papillomatosis-relatied multicystio Lung disease with cidofovir: case report and review of the literature". The cardiopulmonary and critical care journal. The American college of chest physician, 118(4), pp.1210-1214.
5. J.H.Saurat (2000), "Dermatologie et maladies sexuellement transmissibles", Masson, 36me Edition.
6. Kjer J.J, K.EIdQn, Anne Dreisler (1998), "Maternal condylomata and juvenile laryngeal papillomas In their children", Zentralblastt fur gynakologie, Band 110 Heft 2
S, pp.107-110.
7. Padberg B.C. (2002), "Ddtection et typisation des papillomavlms humains en biologie moleculaire pour le depistage des cancers gynecologiques", Fonjm Med Suisse, (42).
8. Yantsos et al (1999), "Recurrent Juvenile-Onset Laryngotracheal Papillomatosis of the Lung", Southern Medical Journal, pp.10130-1015.
IJIMG DUNG KY THUAT RT-PCR TRONG GIAM SAT VA KHAO SAT TINH HINH DICH T I HOC P H A N TUT C U M A/H1N1 TAI MOT SO DON Vj QUAN DOI VIET NAM
N G U Y I N V A N RU*. N G U Y I N THI THANH TAM', N G U Y I N TH| LOAN', LE THU HA*
''** S d m d n Wda sinh - Trvdng Dai hpc DIPTJK Hit N^i
* Vidn VS sinh phdng djch QuSn ddi - Vidt Nam
TOM
TATDS diiu tra dwac Cum A/H1N1 xiy ra d cdc tnr&ng d sw luan chuyen quin sd vi diiu kidn sinh boat khd khan c6 mirc dd nhiim cao nhit ti HQC vl$n H$u Cin, Si quan Luc Quin vi thing 7-9 li th&i diim VIJ dich bOng phit rim rd nhit
Di tii d§ tiin hinh si> dyng kf thudt vi thiit bi RT-PCR giil trinh ti/ doan gen HI cda cdc miu nghiin cOv, cd dd twang ddng cao vi tim ra diim sal khic vdi cic doan gen cOa cic chOng qudc ti. Kit qui chl ra ring cic chOng Iwu hinh tai Vidt Nam khdng chi cd tO- mdt ngudn gdc tir 1 chCing duy nhit mi li cOa nhiiu chOng khic nhau trdn thi gi&i
GUI mS cic doan gen khic cua vims cOm A/H1N1 di phin tich so sinh vdi cic chOng tai Vi$t Nam vi trin thi gi&i de cdcasd dinh gii biin ddi di tmyin vims cOm A/H1N1 hidn nay vi sin xuit vac cin phdng bdnh.
Tip khda: COm ngw&i H3N2 (PB1), COm heo c6~
diin (HI, NP, NS), COm gia dm Bic Mf (PA), COm gia dm Au A, COm A (H2N2),1968: "Dai dich cOm Hdng King", COm A (H3N2)
SUMMARY
Investigated Influenza A/H1N1 cases have occurred in the troop movements and difficult living conditions with the highest levels of contamination institute of Logistics, Army Officers and 7-9 months is the time outbreak most aggmssive
The thread was conducted using techniques and equipment RT-PCR sequencing of the genome of the sample HI, with a high degme of similarity and diffemnces found point to the fragments of the intemational mce. The results indicate that the stmin circulating in Vietnam not only fmm a strain derived fmm only one of the many dlffemnt strains of the worid
Decoding the other gene segments of influenza vims Ai/H1N1 for analysis compamd with strains in Vietnam and in the worid to have an assessment of
genetic variation of influenza vims A/H1N1 and current vaccine production.
Keywords: Influenza A/H1N1, RT-PCR
DAT
VAN D £Nguin g i c virus A (HI N l ) la tai t i hp'p g i m 4 gen cua 4 ehung virus: Gdm gia e l m B i c Mf, Cdm Ipn B i c My, Cum lan chau A va ehdu Au, COm ngu'di.
S y t i hp'p ndy chya cd trydc ddy d ngydi vd Ip-n. V l d u trde vims edm A/H1N1 t h u i e hp: Orthomyxovims.
Kich thu'dc: 80 -100 nm, hinh c l u hodc hinh spi, cd mdng bao lipid. Bd gen: 8 phdn dogn RNA.
D 6 | TUg^NG VA PHUXyNG P H A P L A Y M A U Ddi typ'ng
M l u bdnh p h i m : djeh nhay hpng/tri$u chdng ldm sang dien hinh, nhdng ngu'di tiip xdc g i n
Dan vj
Hpc vidn q u i c phdng, Hpc vidn hdu d n , Hpe vidn Q u i c phdng, Tmng d p thdng tin Hda Binh, Tmdng sTquan lye qudn 1
Dia d i l m
Khoa Vi sinh Y hpc- Vidn VSPD Qudn ddi Khoa Sinh hpc phdn t d - BV Trung yang Qudn ddi 108
D$c diim do-n vj qudn d$l
Dieu kidn s i n g vd sinh hogt tdp trung nhilu ngydi edng 1 thdi d i i m -> tinh trgng Idy nhiim nhanh. Vide phdt hidn nhanh, chinh xac gidp kip thdi d i i phd djeh.
Djch nhdy hpng
Tdch ARN t i n g s i b i n g Kit Qiagen-Ddc rRT-PCR vdi cdp m i i dde hidu edm A: M30F2/08, M264R3/08
Phdt hidn cdm A
rRT-PCR vdi cdp m i i dac hi$u cum A/HI N l SWH1-C0NV-F1, SWHI-CONV-RI Phdt hidn cdm A/H1N1 San p h i m PCR edm A/H1N1
Tinh sgeh san p h i m PCR vdi Kit Bioneer, Fermantas
D l i n di tren gel agarose 2,5% k i i m tra sp PCR,
Y HOC THirC HANH (931) - SO 9/2014
dy dodn Vpcr du'a vdo PCR sequecing PCR cho Sequencing Tinh sach PCR eho Sequencing
Didn di giai trinh ty gen H1 tren mdy CEQ8800 Cdch ldy m l u benh phim vd xd> If
PHUONG PHAP NGHIEN CUU Quy trinh nghidn cdu Liy ifiAu bfnh phim vA xCr 1;^ '
^Thve hl#n phAn ^ng rRT-PCR j , ,^ ^ ChlinjioAn ciim A/H1N1
^. Glil trinh t y gan HI .Xftc,d|nh klAu gen'nguin'g6B^
£x»jF^^g^c*y phi hi^ AftH1N^|j Phyang phdp nhdn gen / PCR
§' , 'W ft - Djch nhay hpng 4S
rRT-PCR vdi c$p moi d^c hif u cum A:
M30F2W8,M264R3/08 _J^
r
Ph^t h i f n cum A [rtCT-PCR vfi-i c^pm6i d|chi|ucCim A/H1N1 Lsj,^.j5WH1-CONV-f1. SWHI-CONV-RI .4,,1 ^ ^ ^ ^ Phdt hi^n cOm A/HI Nl . Phuvng phdp gidi trinh tip / SEQUENCING
Tinh Sf ch sin phim PCR v6i Kit Bioneer,
Fermantas j Difn di tr&n gel agarose 2,5% idem trz $p ^
PCR, d<^ dodn Vpcr dva vao PCR sequecing PCR cho Sequencing Tinh s^ch PCR cho Sequencing Oi^n di gidi Irlnh t y gen HI tren mdy
CEQS800
Phuwng phdp chin dodn phdn ti>
Phyang phap Sinh Hpc Phdn T d TdchcMAt
c^
, L _ _ _ =,- Kitqui
Ky thuat Real-time PCR
- • 1
z S z i
Nguyen ly Real-tanie PCR
K^quA Real-lime PCR Nguydn IJ ky thu^t RT-PCR
3^Sm
Phu'O'ng phdp SANGER gidi trinh t y gen Chu ky sequencing
4 loai ddNTP dypc g i n chit mdu hujnh quang.
Cd 4 phan dng edng xly ra trdn 1 tube cdng di§n di trdn 1 lane.
Mdi dogn AND dygrc ddnh diu huynh quang sd di qua diu dpc d dogn cuii cua gel vd mdu dypc ghi Igi
Y HOC THVC HANH (931) - S6 9/2014
eho tdng ddNTP.
Trinh t y se thu du-pc t d gidi mdu vd m i l mau dgi didn cho 1 logi nucleotide.
Dd dai san p h i m giai trinh t y t i i da Id 700-800 base-pairs.
T e m p l a t e (artglnal) D N A s i r e n d
C3S1
men
unlabeled d N T P s (luorescenlly labollod d d N T P s
za- zsy- zs—
/SV- /Sy-
Z2>- / 5 \ —
G A A A C G G A M^c tieu nghign cOu:
1. X^c <^nh ty 1$ nhi^m cum Aym N1 d mOt s6 (Jon vj quSn doi phia bSc Viet Nam trong vg drcil 2009- 2010
2. YAc S\r\V\ flSc diem itieu gen ma tl6a i<liang nguygn H cOa ciiung vims A/H1N1 so s^nti v6i chung qu6ctS
K£T QUA NGHIEN ciru
1-Klt quS x6t nghidm cum A Ty 16 nhiem cum A
• T$ng»>£tnshKm . T o . a . a d u t n i , n ' n h c u m A
ST quan luc qudn: 82,5%, hpe vidn hdu c i n : 78,57%, hpc vien q u i e phdng (HVQP):70% c i sy thu nhdn qudn td- nhiiu den vj Id y i u t i nguy ca eao cho sy lan t m y i n djch
2- K i t qud xdt nghidm cum A/H1N1 . Ty 10 nliiem ciimA/HlNl
nhci^mA/KlNl
Sy phdn bd phdn tip virus cdm A/H1N1 dgi djch nhieu han so vd'i phdn tip cdm A cdn Igi cho thiy md'c dd Idy lan eua vims ndy. Ngodi vims A/H1N1 edn ed H3N2, H5N1 eho t h i y nauydn nhan phd-e tgp cda cae tridu chd'ng Idm sdng n l u ed d i n g nhilm hay tdi t i hgp
3- Phdn bd cum A/HI N1 theo thdng T y l ^ n h i e m c u m A / H l N l t l t e o t h i ^
Giai doan nhidm cdm A/H1N1 cao nhlt Id td- thdng 7 den thdng 9
4- K i t qua Realtime RT-PCR
1 2 3 4 5 6 7 8 9 1011 12 13
I C D m A M I : 349bp '
6 7 8 9 10 11 12 13 5- Kdt qud gidi trinh X\r virus cdm A / H1N1, VietNam 05/2009
rslWil\
-/U.-Ay'WvwV- -K ,'VAA/^./>fU^
•J-^h
V HOC THVC H A N H (931) - SO 9/2014
6- K i t qua Gidi trinh t y nucleotid cda virus 9- K i t qud t i m su> s a i khdc acid a m i n t r o n g cdu trOc protein cda v i r u s
8- Kdt q u d t i m sv s a i khdc nucleotid t r o n g phdn t£r acid nucleic
QiBfysfqiEticMlMlSMGilGCaATTGSSCICiGTGTCHTCATnGAMEGTITGASSTATTCCCAiiJLGSCAftG 9|n6937136|^|l71(3»)SGAGSeCiriTTGBGCTCiGlGIC'iTCATnGiS*eem5M.imTCCC»AGBQRO icimi
7- K i t qua BLAST len ngdn hdng gen thd gid-i NCBI
10- Kdt qud xdc d j n h d o t bidn SNP trong cdu t r d e b i i n t h d cua v i r u s c u m A H1N1 V i ^ t Nam
iniswnrriiiniiihRniii'i.iiniii.iii'iiinnnninr im
iiiiii iiMir niiTinAi/iiiiiii iiiiiiii-i I luiiiid iiiiiiiin. • jHiiii 1 iiiniiii ii'ii iiBb n ciuniCNuiii Id Li'iriiiiiiiiiiiiiiHi Ill I mil luiii'iMiiRn ILLUil'l I
Uimiiiii' iiAi/liili LI ilili I IMIIlilllMLI'liriii
^Nm:iWlil.Bl'lMii'. I II 11 Ml I I n ur
•iiiluwiil iiriiiiwiiiiii I • I'll I Ml 11 uiiiiiiyL iiiiiijR iM I.I.I III' iiii I I'l uiii ••• 'I nil 111 HiiiiiiHr milLIR III 11 i l .l_l I I I ili'lllil I • 11 l-l M nil liBL .inuuv >ii 11 Mill' .1.111 Ki'ii 'I • I iiiiiiLiii. niiiiiiii.
•iniuiii III 111 I'l- nil 11 iiniii-ii >i III iir nni iiui iiinii'ii III 1.1.111111 iiii 11> iiiiii ill 'h'\ I III 1 iiiiiiu . 'iniiiB III 11 ii'ii 1.111. iii.'i ii.>i in III • iiiiiiiB IIHIiMi|||.l.l.liii(i li'i 1 illfii i. I llil.lli,illlliiilK 'INiliR >llil.l.>l III .III ,1 >1l'l 11 11II ill. nilJIIIBL JinHiR'-ll LIIHilU li II II • III'IILI 111 III IdJIIMMIU iiniiiiiii»i.iii.ii "111.1 iii>i niiiii iii<i liiin.i'niiii'iM. >
K.... .,..-.,,, ^,|„|„ 1,1,111,11 , ^ „||||,|,y
•iiiiiiai iiiiiiiiiiMiiiiLiii>i •wiiiiiiiii.inii'i I u'liiiiRL llliHikll.lHIII!IU.IIilli1.ilif.l|i|lililK
B A N L U A N
1 . K i t qud g i d m s d t ca b^nh n h i d m virus ciim A/H1N1 v d t $ 1$ ngyd>l Idnh mang mdm b$nii khdng tridu chipng t a i cdc 6cm v i q u d n ddi midn Bde td-2009-2011.
K i thudt realtime RT-PCR TaqMan Probe phdt hidn d i u I n cdm A vd edm A/H1N1 dd dype dng dung thdnh edng vd'i dd ddc hidu cao (khdng cho phdn irng d y y n g tinh gid vd'i virus edm B vd cdm A/H5N1) vd ngyd'ng phdt hidn Id 102 copies/ml.
Vide d-ng dyng kT thudt realtime RT-PCR trong gidm sdt dich cdm A/H1N1 dgi djch ndm 2009 -2010 tai b i n den vj qudn d i i : Hpc vidn Hdu c i n , Hpc vi§n q u i c phdng, Tryd'ng Tmng d p thdng tin, Tryd'ng sT quan lye qudn dd riA ra d u v c mdt s i nhdn xdt nhy
Y HOC THVC HANH (931) - s6 9/2014
cac ca bdnh d i u tidn du'pc xac djnh dycng tinh vdi cdm A / H I N I h l u h i t cd trieu chd-ng iam sdng giong nhau vd deu a t h i cdm nhe. Chya ghi nhdn dyp'c tryd'ng hp'p tu- vong ndo cd xet nghi0m dyong tinh vd'i cdm A/H1N1.
Djch x l y ra rdi rdc o- t i t ea ede thang trong ndm, tuy nhidn s i xdt nghidm du-ong tinh eao nhlt Id vdo cde thdng td- 7 d i n 9.
Nguydn nhdn lan t m y i n djch bdnh tren qui md I6n v i khdng gjan vd thd'i gian chd yeu do m i t s i dde (Jem djeh td nhu-: ed s y di chuyen qudn vd thu nhan qudn v&] s i lu'gng Id'n td* cdc khu vyc dang cd djeh lyu hdnh; do'n vj ddng qudn tai vdng dang xay ra dich.
Yeu t i d i l u kidn s i n g chat hep va d i i u kidn vd sinh mdi tnj'dng doanh trai chu'a that s y Id nguydn nhdn chinh d i n den kha ndng bung phdt dich bdnh.
Hlu h i t cdc ca benh cd xdt nghiem du'ang tinh d i u khflng cd t i l p xde trye t i i p vd'i ngu'd'i dang nhiim cdmA/H1N1.
Djch bdnh ed kha ndng d i i n b i i n t i t hen khi ed s y can Uiiip dung edeh v i phdng ehing tai cdc don vj sau khi drnp^c edng b i djeh.
2. K i t qud khao sdt d$c d i i m sinh hpc phdn tu>
vims cdm A/H1N1 tgi cdc d e n vj qudn d^i khu vyc m i i n B l e tCi* 2009-2011.
Hodn toan giing dac d i l m virus cdm A, sy t i i n hda thing qua cdc ^ a y d i i trong cau trde phdn td' se dypc b i l u hipn bang s y t i l n hda ve khdng nguydn, thay d i i ddc tinh khdng nguyen vd "trin thodt' p h i midn dich dd cd trong q u i n the ngu'd'i. Vi vdy kiem sodt sy thay doi phdn tu* eda vims cum Id edng vide thydng xuydn trong gidm sdt virus edm. D l tdi mang lai nhO'ng thdng tin vd nghidn cd'u s y lyu hdnh cda cdc vims edm theo mda A/HI N l .
K£T LUAN
1. Cdm A/H1N1 xay ra d' cdc tru'd'ng cd s y ludn chuyin qudn s i vd didu kidn sinh hoat kdm Id cao nhlt: HVHC, SQLQ. Thdng 7-9 Id thd^i d i l m vy djeh bdng phdt r i m r i n h l t
2. Dd giai trinh t y dogn gen H I cda cde mau nghidn cd'u, ed dd tu'O'ng d i n g eao vd tim ra (Siro sai khde vd'i cdc doan gen eda cdc chdng q u i e t l . cae chdng lyu hanh tai Viet Nam khdng chf cd td* mdt nguin g i c td' 1 chdng duy nhlt ma la cda nhiiu chdng khac nhau trdn t h i gid'i
3. €)d su- dung ky thudt Realtime-PCR d i xdc djnh
cac tru'd'ng hp'p nhiim edm A/H1N1 td- bdnh p h i m djeh ngody hpng eda bdnh nhan nghi nhilm edm A/H1N1. Tach chilt ARN, phien ma ngyp'c, ehu trinh PCR dk dinh tinh. Thu s i n pham t i i n hdnh thyc hidn giai trinh t y gen.
Dd xdc djnh du'p'c ty le nhiem edm A/H1N1 vd t f Id ngyd'j Idnii mang bdnh tren mdt s i dan vj qudn d i i phia B i c Vidt Nam.
4. Dd thyc nghidm giai trinh t y 20 mdu edm A/H1N1 d i xde djnh gen ma hoa td' dd BLAST qua ngdn hdng gen tim s y tu'ang d i n g vd'i ede ehung virus cdm A/HI Nl dd biit tren t h i gid'i.
5. Oa xay dyng cdy pha hd eho cdc chdng virus d i i n hinh thyc nghidm, d i d y a ra djch t§ hpe phan td- vims cum A/H1N1 trdn nhyng don vj l i y m l u benh p h i m , xde i^nh djch t i hpc phan td-
Dd xdc djnh cdc d i l m d i t bien cda vims cum lyu hanh tgi Vidt Nam so vd'i chdng virus cdm A/H1N1 dd du'pc xac dinh trdn t h i gid'i cd s y tyong d i n g cao nhlt.
TAI LIEU THAIVI K H A O
1. B i Y T i (2009) quyet ^ n h v l vide ban hdnh hyd'ng d i n k ; thudt l i y mdu, tiao qudn, vdn ehuyin m l u bdnh pham cdm A(H1N1)
2. B i Y Te (2009) Hu'd'ng ddn c h i n dodn, d i i u trj vd phdng ldy nhidm edmA(H1N1).
3. N Engl J Med 22 May (2009), Novel Svi/ine- Origin Influenza A (H1N1) Vims Investigation Team.
Emergence of a Novel Swine-Origin lnfluer\za A (H1N1) Vims in Humans. World Health Organization (WHO). CDC protocol of realtime RT-PCR for Influenza A (H1N1). 30 April 2009.
4. The Domestic and Intemational Impacts of the 2009-H1N1 Influenza A Pandemic. Institute of Medicine (US) Fomm on Microbial Threats.
Washington (DC): National Academies Press (US);
2010.
5. Evaluation of four realtime PCR assays for detection of Influenza'A {H1N1)Viruses.
Eurosurveillanee, Volume 14, Issue 22, 04 June 2009. Sultan Qaboos Univ Med J. 2010 August;
10(2): 187-195.
6. S. J. Flint. L W . Enquist, V. R. Raeaniello, and A. M Skalka. 2004. Principles of Virology: Molecular Biology, Pathologu, and contml of animal Vimses.
Second Edition. ASM Press Washington, D.C.
THUC TRANG TAI C H I N H TAI CAC BENH VIEN DIEU DUtlNG - PHUC HOI CHUNG NANG
TOIVITAT
B^t vin de: Tii chinh y t4 li bd phin chinh ciu thanh hf thdng y t4 quSc gia.Trong bdi dnh hing lo^t cic chinh sich vd tw chO tii chinh cdng mdi tfupc ban hinh, vdn dd dinh gii tinh hinh tii chinh
N G U Y i N TH! M1''H T H O Y , Bai hgc Ytd Cdng cong TRAN QUV TLtdNG, Bd Ytd cic bdnh vien didu du&ng phuc fidi chOc ning (BVBDPHCN) ii rit cip thiit Muc tidu: Md ti thuc tr$ng tii chinh tai cic BVDDPHCN giai dOin 2007- 2009. Phuvng phip: Md ti cSt ngang trdn 35 BVDDPHCN trong ci nutic. Kdt qui: Kinh phi cip
