Y HOC VI&T NAM THANG 6 - S01/2?
Ruggeri E.M, Carlini P, Poilera C F et al (2005), " Long tenn survival in locally advanced oral cavity cancer: an analysis of patients treated with neoadjuvant dsplatin-based chemotJierapy followed by surger/', Head neck 27(6), pp 452-458.
Pham Tuan (1987), "Dieu trj ung thu" luQi d gia doan muon bang hoa chat dong mach ket hdp vd tia xa", torn tat cong trinh nghien cuU khoa ha ung 'thW1987, tr 135-136.
NGHIEN CU'U U'NG DUNG KY THUAJ ELISA PHAT HIEN DOC TO VI KHUAN T A
Ph^m Dire Minh*, Nguyin Hung Long**, Hoang Van Lvtrng*
T6|V1 TAT
NhiSm vi khuan ta ho^c dpc ^ vi khuan ta se gay tieu chay c3ip va nhOhg hau qua nSng ne tuy theo kha nSng mien djch ciia cd the vat chu. Trong nghien cijtj nay, chung toi \Jnq dung khang the ddn dong de thiet ke phan iihg EUSA nham phat hien dpc to CTX ciia vi khuan ta, ilhg dung tmng chan doan, dieu trj va di/
phdng. Ket qua nghien ciTu cho thay phan utig ELISA CO kha nSng phdt hi§n dude ddc to d nong dp lOng/mL Cac khang the co dp nhay khac nhau trong phat hi?n CTX, theo thuf ty* co do nhay tir thap ten cao 1^: C01238M, C01592M, C01593M, Cdl594M.
TO" khda: Ooc to ta, Khang t h i khang CTX; Vi khuan ta; EUSA
SUIVIMARY
APPLIED RESEARCH ELISA T O DETECT CHOLERA T O X I N
Infection with Vibrio cholerae or cholera toxin causes acute diarrhea and severe consequences depending on the immunity of the host. In this study, we applied monoclonal antibody at ELISA, designed to detect toxins CTX of Vibrio cholerae, applications in diagnosis, treatment and prevention. The research results show that EUSA is capable of detecting toxins at concentrations of lOng/mL. These antibodies have different sensitivities in detecting CTX, in order of sensitivity from low to high are: C01238M, C01592M, C01593l^, C01594M,
Key Y/ords: CTX. monoclonal antibody to CTX;
Vibrio cholerae; EUSA.
I. OAT VAN o l
Tieu chay cap do ta ta benh truyen nhiem cap tfnh do nhiem vi khuan t^ du'dng tieu hoa.
* Hqc vt$n Quan y
** Cue An toan Ihyt Pham -BoYte Phan bi$n Ichoa hpc: PGS.TS. Ph^m Ngpc Chau
Benh cd the gay ra cac vy djch Idn va du'dc col la mpt benh truyen nhiem nguy hiem, do klia nang lan truyen nhanh trong cpng dong thdng qua thirc an va/hoac nguon nu'dc sinh hoat 6 nhieni bdi vi khuan ta (VKT). Sau khi nhiem VICT theo du'dng tieu hoa, dpc to do vi khuan tiet ra se bam va tham nhap vao te bao niem mac ruot, gay roi loan qua trinh van chuyen ti'ch ciTc cac chat dien glai qua mang te bao niem mac mot, dan den tang tiet dich vao long ong tieu hpa va gay tieu chay mat nu'dc [ 3 ] , [ 4 ] . Phat hien nhanli dpc to vi khuan ta trong thiTc pham va mpt s6 benh pham khac se giiip cho qua trinh chan doan, dieu tri va du" phong dat ket qua cao [1], [ 2 ] . Oe tai du'dc tien hanh nham thiet ke ptian uhg EUSA va tim ra nhuTig khang the toi iTu ntiat de phat hien dpc t d CJY, bang phudng pfiap EUSA cung nhu' img dung tren cac phan ling mien djch khac.
I I . 0 6 l T i r O N G , VJSiT U £ U VA PHUaNG PHAP NGHIEN CO'U
2 . 1 . D o i tu'dng, v a t llSu nghiSn c(hi - Ooc to CVK (G-117 Lot # P5351) ciia cong ly Enzo Life Sciences
- Hoa chat, sinh pham chu yeu; khang the khang CTX cua hang r^eridian, cpng hdp peroxidase • khang the IgG ciru khang chupt va cac hpa chat can thiet khac cho phan ii'ng EUSA (Bio-Rad, Hoa Ky; Sanofi, Phap),
2 . 2 . Phu'dng phdp n g h i e n cdu
* Chuan bi khang nguyen: khang nguyen du'dc pha loang ra nhieu nong dp khac nhau ti danh gia
* Phan Ong EUSA phat hien dpc to CU (thu'c hien tai La bo Mien djch, Trung tSm Ngtiifij ciru Y - Du'dc hpc quan sir, Hpc vien Quan y):
46
,icT HAM THANG 6 - s61/2014 Quy trinh xet nghiim EUSA phat hiin
a^ to CTX;
• BVdc 1: chuan bj bo EUSA iam xet nghiem g3n tren gia dd.
• Bu'dc 2: cho 100 ul mau thiJ (dpc to CTX d cac nong do Ithac nhau) vao moi gieng, 2 gieng doi chulig cho dung djch i<hang nguyen chuan, chijng am. U 4°C qua dem.
• Birdc 3: riia 5 lan bang dung dich riia PBS-Tween.
' Bu'dc 4: Blocking bang sSa (5% sl<im milk/PBS Ix) / 3h 6 nhiet dp phong; 300 ul/gieng
• BUdc 5: Lip lai bu'dc ru^ nhu' bu'dc 3.
• Bu'dc 6: dita khan^ the de khang chuot die hieu cn vao cac gieng. U 37X tnjng 15 phut.
• Bu'dc 7: Lap lai bu'dc riia nhu' bu'dc 3.
III. KET QUA NGHIEN CO'U
3.1. Kha nSng phat hien khang nguyen CTX cua phan ulig ELISA su* dung khdng the
• Bu'dc 8: them khang the khang IgG de gan HRP vao cac gieng. U 37°C trong 15 phut.
• BUdc 9: Lap iai bu'dc riia nhu bu'dc 3.
• Bu'dc 10: them cd chat mau. I^oi gieng 100 ni cd chat TMB nong dp 0,5 mg/ml bo sung them 0,006% H2O2. Ooc ket qua sau 5 phut, quan sat sir chuyen mau cua cd chat tlT khong mau chuyen sang xanh iuc bang mat thu'dng, them vao a xit sulfuric de doi mau xanh iuc sang vang va do mat dp quang hpc d bu'dc song 450 nm bang he thong DTX 880 (Beckman Coulter, My).
ThSng ke vi so sanh ni$t do quang hpc ciJa cac gieng ti/dng iJng vdi cac mau huyet thanh bang cac thuat tpan cua phan mem l*licrpsft excel 2003.
Nh$n x « : Gia trj OD trung binh cua gieng am + 2SD la khoang 0,05. Qua ket qua thi nghi?m cho thSy khing the C01238l>1 ijng dung trong phan uTig ELISA cd the phat hien dtfcjc khing nguySn CTX. d n6ng dp lOng/mL. Kha nSng phat hien tSng khi nong do khang nguyen tang. Tin hieu ro net d nong khang nguyen 200ng/mL. Khang the C01238I"! cd dp dSc hieu tuyet dpi vdi khang nguyen CTX.
3.2. m>i ning phM hl|n khing nguyen CTX cua phan Mng ELISA s^ dung khing t h i Cai592M
Nh$n xit: Qua ket qui thi nghl$m chP thay khing the C01592M litig dung trong phan utig EUSA cd
the phit hl^n dm?c khang nguyin CTX d nong do lOng/mL. Kha ning phat hl?n tSng khi nSng dp
khing nguyin tSng. Tin hiiu ro net d nong khing nguyin lOOng/mL. Khing the C01592M c6 dp die
hiiu tuy$t dol vdl khing nguyen CTX.
Y Hpc VIET NAM THANG 6 - S01/201
3.3. Kha nang phat hien khang nguyen CTX cua phan irng ELISA siJf dung khang the C01593M
Phan i>nQ Et-ISA ptiat hi^n CTX b a n g khang thS COI 593M
Nhan xet; Qua kit qua thi nghiem cho thay khang the C01593l«l irng dung trong phan uTig EUSAco the'phit hiin du'dc khing nguyen CTX d npng dp lOng/mL. Khi nang phit h]en tang khi npng d?
khang nguyin tarig. Tin hiiu rp net d nong khang nguyin 80ng/mL. Khang the C01593M cd dp d3c hieu tuyet dpi vdi khing nguyen CTX.
3.4. Kha ning phat hien khang nguyen CTX cua phan u'ng ELISA sU dung khing tlijl C01594M
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Nh$n xit: Qua kit qui thi nghiim chp thay khing the C01594M irng dung trpng phin u'ng EUSA eJ the phit hiin du'dc khing nguyin cn d npng do lOng/mL. Kha nang phat hien tang khi nong dj khing nguyin tang. Tin hiiu rp net d nong khing nguyen 60ng/mL. Khang the C01594M cp dp iljc hiiu tuyet dpi vdi khing nguyen CTX.
IV. BAN LUAN
Phan ling ELISA la mpt trong nhiilig phin utig mien djch cp dp nhay cap nhat. Trpng nghiin ciru liay, ELISA du'dc diing de danh gia khi nSnp phit hien cua mpt sp khang the ddn dong dpi vdi khiiig nguyen CTH cua ddc tp ta [6], [7]. Kit gua chp thay phan ijng EUSA su' dung ddn khang the cp the phit hiin khang nguyin CTX d nong dp lOng/mL. Cac khang the CO kha nSng phat hiin khing nguyen khic nhau, xep theo thu' bi tang dan cua do nhay lan lu'dt la cic khing the C01238M, C01592M, C01593M, C01594M.
Do cac khang the nay deu la khang the d ^ clone va cd vj tri phit hien khang nguyen r hieu (epitope) khic nhau (theo thong tin cua r sin xuat) nin hoan toan co the ifng dgng gra cap khang the trong cic phan iTng mien dich k'' sandwich t2J. Ket qua nghiin cuXi nay s i la ( cif de cd the CP cic ii/a chpn hdp ly tiiy thep y cau cu the ciia cac u'ng durig khac nhau. .
Tai Viet Nam, vi khuan ta van du'dc phit hii nhutig chira gay djch. Viic thiit k i cpng cy ffl sin sang phat hiin va phpng chpng la rit dil thiet [5].
. NAM THANG 6 • sd 1/2014
\i. K f r LUAN
Da thiet tte thanh cong phan iTng mien djch ELISA sir dyng^ l<hang the ddn clone de phat hien 3pc to vi Ithuan ta. NguSng phat hien ciia phiin irng la lOng/mL.
Cac Ichang the co kha nang phat hien CTX khac nhau, theo thir tu* khang the co do rihay tir thap ten cao fan ti/dt ta: C01238M, C01592l^, C01593l^, C01594M.
T A I L I | U THAIVI KHAO
1. Phiing Dac Cam, 2003. Vlbho cholerae va b&Jfi dich ta. NXB Y hoc.
z. Do KhSc D^i/ Nguyen Tnfdng Sdn^ Le V3n Ddng, 2011. IVghien aitj che tao xet nghiim EUSA kieu sandwldi sii" dung 3 bai khang the d$c hi$u phat hien noc rSn luc xanh va ho dat
Tap d l i y di/dc hoc quan sir 2011, so 9 - 2011, Tr. 11-19.
3. Tr^n Linh Thu'dc, 2002. Phudng phap phan tich vi sinh vat trong nu&c, thut pham va moi tru&ng. NXB Giao due.
4. CDC. 1999. Laboratory Methods for die Diagnosis of Epidemic Dysentery and Cholera.
5. CDC. 2003. Agents for bioterrorism.
6. Huu Dat Tran, Munirul Alam, Nguyen Vu Trung. Multi-drug resistant Vibrio clioierae 01 variant El Tor isolated in northern Vietnam between 2007 and 2010. J Med Microbiol. 2012, 61(Pt3), pp.431-437.
7. Kazuyuki H, HideyukI A, Koji U. Passive oral immunization by egg yolk immunoglobulin (IgY) to Vibrio cholerae effectively prevents cholera. Acta Medica Okayama. 2010, 64 (3), pp.163-170.
NHAN XET Sir THAY DOI HUYET BONG SAU AP DUNG LIEU PHAP DIEU TRI SOM THEO MUC TIEU 6^ B I N H NHAN SOC N H I I M KHUAN
T 6 M TAT
Soc nhiem khuan (SNK) id m0t benh ly thu'dng g^p trong hoi SLTC cap cu'u c6 ti l | tir vong cao. Ll^u pliSp <liBu trj sdm theo muc tieu (EGDT: early goal directed therapy) la chien \\i(}c dieu trj se giup cai thl^n b' l | tir vong, ngay nSm vien vd chi phi dieu tr;.
Nghign alU nSy d^nh gid si/ thay doi huyel dgng sau dp dgng EGDT t benh nhan SNK. Phu'dng phap: 32
! b§nh nhSn vdl chan dodn SNK du^c dieu trj tich cue I trong 6h dSu vdo vien vdl d$t catheter tTnh mach trung I tdm, hfil silt dich duy tri CVP 11-14 mmHg, HATB >65 mmHg, He >30%, Scv02 £70%. Sau do cac b^nh nhdn ndy du'(?c chia lam 2 nhdm thanh cong hodc that b^l vd tigp tyc theo doi cdc dien bien huyet ddng bao
\ gSm CVP, HATB, Scv02, acid lacdc sau dieu tri. Ket
; qud: Sau dp dyng EGDT, cdc chi so huyet dpng cdi ' thl?n cd ^ nghTa thong ke sau 6h dieu hi. Nhom tiidnh cong c6 tl I? tudi mdu to chilt tot hdn c6 y nghia thdng ' k§ so vdi nhdm that b?i vdl p<0,05. KSt lu$n: U^u ' phdp EGDT gliip cai thien cd y nghTa cdc chi so huygt ' d$ng vd cdc chi so oxy hod to chut d benh nhdn SNK.
TCiithoi: Ueu phap dieu tri sdm theo muc tieu,
• SNK, Scv02.
* B$nh vl?n B^ch Mai
"B^h vl$n ThalNguy^
Phin bl$n Ichoa hpc: PGS.TS B§ H3ng Thu
Nguyen Hfru Quan*, Mai Duy Ton*, Vu Hai Yen**, Le Thj Diem Tuyet*
SUIVIMARY
TO EVALUATE THE EARLY GOAL DIRECTED THERAPY (EGDT) EFFECTIVENESS ON HEMODYNAMICS A N D TISSUE PERFUSION
I N D E X I N S E P n C SHOCK PATIENTS Background: Septic shock is common in ICU and has high mortality. EGDT has been used early to help Improve survival rate and reduce complication. This study was to evaluate the EGDT's effectiveness on hemodynamic and tissue perfusion improvement including pulse rate, mean arterial pressure MAP, CVP, Scv02 and plasma lactate. Method: There were 32 patients with septic shock enrolled and were resussitated with EGDT on the first 6 hours after admission. These patient were then divided into two groups; NI success 22 pts (68,75%) and N2 feilure 10 pts (31,25%). Results: In septic shock, There were significantiy improvement in hemodynamic parameters and tissue perfusion index (p<0,05) after the first 6 hours. There was significantiy difference between NI success group and N2 ftilure group about Scv02 and plasma lactate after Implementing EGDT Conclusions:
EGDT hepis impove the hemodynamk: parameter and tissue perfusion index in septic shock patients.
Keywords: septic shock, EGDT, 6 hours
