TCNCYH PhtJ trWdng 62 (3) - 2009
TIM HIEU GIA TRj CUA PCR - BK TRONG ^
CHAN D O A N T R A N D I C H M A N G PHOI DO LAO
Pham Thj My Dung^ , Tran Hoang Thanh^
^Sinh vien Dai hoc Y Ha Ngi
^Dai hoc Y Ha Ngi
Muc tieu: (l)Tlm hieu gia trj cua Polymarase chain reaction - Bacillus de Kok (PCR - BK) trong chan doan .tran djch ming phdi (TDMP) do lao. (2) Xic djnh mdi lien quan gida PCR - BK vdi mdt sd bleu hien lam sang vi can lam sing cua TDMP do lao. Bdi tugng va pbuang pbap ngbien cdu: hoi cdu 127 BN dugc chin doan TDMP do lao dugc dieu trj ndi trd tai khoa Hd hap - bdnh vien Bach Mai. Ket qua: 32,3%o trudng hgp cd PCR - BK djch ming phdi duang tinh. Khdng cd mdi Hen quan nio gida PCR - BK vdi sdt, tdc do miu ling, phan dng Mantoux vi thdi gian khdi phit bdnh
Tir kboa: tran dich mang phdi do lao. Polymerase chain reaction - Bacillus de Kok (PCR - BK)
I. DAT V A N DE
T r i n djch mang phdi la mot hdi chflng thfldng gap trong thflc hanh lam sang do nhieu nguyen n h i n , trong dd do lao la mdt nguyen nhan thfldng gap vdi ty le kha cao d mpi cd sd Y t l tfl trung fldng den dia phfldng. De c h i n d o l n x l c djnh tran djch mang phdi (TDMP) do lao, sinh t h i i t m i n g phdi I I mdt ky thuat td'i flu, nhflng cdn cd han c h l n h i t dinh do chi ap dung dfldc trong d c trung tam y te' chuyen sau. Do vay muc tieu cua de t i i nay la:
1. Nghien cufu gia tri ciia PCR - BK trong chan doan TDMP do lao.
2. Xac djnh mdi lien quan giifa PCR - BK vdi mgt so bieu hien lam sang va can lam sang cua TDMP do lao.
II. DOI TUONG VA PHU'ONG PHAP NGHIEN CL/U
1. Ddi tfldng nghien cu'u
Tieu chuan chgn benh nhan
- C I c benh nhan dflpc chan doan xac djnh TDMP do lao vdi cac tieu chuan vang hoac tim t h i y AFB trong dich m i n g phdi hoac tdn thflong lao trong md benh hpc cua sinh t b i l t mang phdi.
- Tat d cac dd'i tfldng nghien cflu d i u dfldc l l m xet nghiem PCR - BK djch mang phdi tai vien Ve sinh Dich te Trung fldng. Ky thuat dfldc tie'n hanh nhfl sau:
- Benh pham dflpc l i y vd khuan vdi khoang 5 ml, dfldc dflng vao d'ng nghiem vd trung mdi (de tranh g l y dfldng tfnh g i l ) . Bao quan benh p h i m d nhiet dp - 20°C - 4°C va khdng nen gifl > 2 tuan n i u chfla cd d i i u kien sfl dung.
- Benh pham thfldng dflpc xfl ly theo phfldng p h i p Boom: dung guanidium thiocynat - GuSCN de ly giai te b i o v l b i t boat man nuclease, bdc Id A D N va t i c h Ipc chung bang each gan len d c hat silica hoac diatum.
- Chuan bi hdn hpp p h l n flng gdm 4 loai:
nucleotid (2mM), 2 doan A D N mdi (0,2 |jm/mdi doan), men polymerase (1 V/50 microlit trong dung dich dem TrisHCL (10 M m TrisHCL va 50 m M NaCl) cd chfla M g C b (2 mM), uracil A D N glycosylat (UNG) dflpc cho them vao hdn, djch (0,1 V/50 |jl) de chd'ng nhiem.
- Phan flng dfldc thflc hien qua 40 chu ky va mdi chu ky gdm 3 giai doan:
-I- Giai doan b i i n tfnh: 94°C trong 15 phut.
-t- Giai doan lai ghep: 65°C trong 2 phflt.
TCNCYH Phu trWdng 62 (3) - 2009
+ Giai doan tdng hpp: 72 C trong 3 phut - Tien hanh dien di de p h l t hien san pham PCR tren gel agarose 2% trong dem TBE (Tris 0,89M; acid boric 0,89M; EDTA 0,02M) vdi cfldng dp ddng dien 2 - 3 vol/cm^) chup I n h gel bang tia huynh quang.
D i n h g i l k i t q u i : do PCR cd dp nhay cao (cd the phat-hien dflpc 10 fg (tflPng dflpng 1 - 3 vi khuan trd len) nen ket qua can dfldc phan tich, d i n h g i l so sinh vdi chflng am (chi cd dung djch hdn hdp phln flng) va chflng dfldng (ADN cua M. tuberculosis).
I I L K E T Q U A
1. Phan bo' benh nhan theo tudi
Tieu chuan loai trd
- Nhflng benh n h i n khdng cd mdt trong d c tieu chuan vang neu tren.
- Nhflng ddi tfldng khdng dfldc lam PCR - BK trong dich mang phdi.
2. Phfldng phap nghien cu'u Hoi cflu md t l e l t ngang.
Thu thap so lieu theo hd sd v i o benh I n m l u . Xfl ly sd lieu bang p h l n m i m SPSS1 5.0.
\ 1 . tl
Bieu dd 1. Pban bd benb nhan theo tudi (n = 124)
Nhan xet: nhdm tudi < 40 chiem Uu the ban mgt ntfa sd benh nhan (52.yi°/o) thugc nhdm < 40 tudi, trong sd niy cd 55,8%o d nhdm tudi td 20 - 29 tudi.
2. Phan bdbenh theo gidi (n = 127): nam gidi c h i i m dai da sd(nam: 66%; nfl: 34%. Ty le nam/nfl: 2/1).
3. Phan bd theo nghe nghiep (n = 127): d n bd cdng chflc, trf thflc c b i l m flu the vdi 60,7%. Ndng d i n vdi ty le t h i p hpn vdi 2 9 , 1 % . '''
4. Tinh chat khdi phat cua b?nh (n = 127): benh khdi p h l t c i p tinh chiem ty le kha phd bien (Cip:
58,5%; b i n c i p : 18,9% va man tfnh: 12,5%).
5. Ke't qua phan u'ng Mantoux (n = 81): 38/81 (46,9%) trfldng hpp cd p h l n flng Mantoux dfldng tinh [ndt s i n 5 mm (32,1%); 5 - 9 mm (21%); 10 - 15 mm (32,1%); 16 - 20 mm (8,6%); > 20 m m (6,2%).
TCNCYH PhtJ trWdng 62 (3) - 2009
6. Ke't qua PCR - BK djch mang phdi
Bang 1. Ket qua PCR - BK dich mang phdi (n = 127) P C R - B K
Dfldng tfnh A m tfnh
So' ngfldi 41 86
%
(32,3) (57,7) Nhan xet: 1/3 ddi tdang nghien cdu cd PCR - BK djch mang phdi ditang tinh.
7. Lien quan giu'a PCR - BK djch mang phdi vdi sd't (n = 124)
Nhan xet: khdng cd mdi lien quan gifla PCR - BK dich m i n g phdi vdi sd't 8. Lien quan giu'a PCR - BK dich mang phdi va tinh chat khdi phat benh
Bieu do 2. Lien quan gida PCR - BK dich mang phdi va tinb chit kbdi phat benb (n = 127) Nhan xet: Khdng cd mdi lien quan giita PCR - BK djch mang phdi vi tinh cha't khdi phat benh.
9. Lien quan giu'a PCR - BK djch mang phdi va phan u'ng Mantoux
Bieu do 3. Lien quan giifa PCR - BK dich mang phdi va pban dng Mantoux (n = 81) Nhan xet: khong cd mdi lien quan giita PCR - BK djch ming phdi vi phan t'fng Mantoux
TCNCYH PhtJ trWdng 62 (3) - 2009
10. Lien quan giifa PCR - BK dich mang phdi vdi tdc do ldng mau (n = 78)
Nhan xet: khdng cd md'i lien quan gifla PCR - BK djch mang phdi vdi td'c dp l i n g mau.
IV. BAN LUAN
1. Tuo'i: hpn phan nfla (52,2%) sd dd'i trflpng nghien cflu cua chung tdi cd dp tudi < 40. K i t q u i nghien cufu cua chung tdi cao hon mot chut so vdi ket qua nghien cflu cua Dd C h i u Hung [2] vdi 46,34%.
2. Gidi: nam gidi la dd'i tflpng hay gap n h i t vdi 56,6% trong cac dd'i tfldng nghien cflu cua chung tdi. K i t q u i nghien cflu nay cung tfldng tfl nhfl k i t q u i nghien cflu cua Dang HiJng M i n h [3] vdi 5 8 % la nam gidi.
3. Nghe nghi?p: c l n bd va trf thflc la hai dd'i tflpng chung tdi hay gap n h i t . Khac biet vdi nghien cflu cua Le Thanh Chfldng [1] gap chu yeu la ndng dan. De g i i i thich sfl k h l c biet n l y chung tdi cho rang ly do chfnh la do gan day sfl hieu b i l t v l t r i n djch mang phdi tfldng ddi td't hdn dd'i vdi hai dd'i tfldng nay nen hp d i n gap thay thud'c sdm hpn.
4. Tinh chat khdi phat cua benh: 68,5% so dd'i tflpng nghien cflu cua chung tdi cd khd phat benh c i p tinh. K i t q u i nghien cflu cua chung tdi cao hon k i t q u i nghien cflu cua Nguyen X u I n T r i l u [5], nhflng khdng n b i l u vdi ty le gap 5 7 % .
5. PCR - BK dich mang phd'i: chung tdi chi tim dfldc PCR - BK trong djch mang phdi dfldng tfnh vdi 32,3%. K i t qua cua chung tdi tfldng tfl k i t qua nghien cflu cua Au Thanh Tung [5] vdi ty le 35,5% va cua Blanie M (2005) vdi ty le 3 2 % .
6. Cac mdi lien quan giiJa PCR - BK vdi mot sd thdng so khac
- Khdng cd md'i lien quan gifla PCR - BK djch mang phdi vdi trieu chflng sd't.
- Khdng cd md'i lien quan gifla PCR - BK djch mang phdi vdi tfnh c h i t khd p h l t cua benh.
- Khdng cd md'i lien quan giOa PCR - BK dich mang phdi vdi p h l n flng Mantoux.
- Khdng cd md'i lien quan gifla PCR - BK dich mang phdi vdi td'c dp lang mau.
V. KET LUAN
1. 32,3% trfldng hdp PCR - BK djch m i n g phdi dfldng tfnh trong d c benh n h i n t r i n djch mang phdi do lao.
2. Khdng cd md'i lien quan gifla PCR - BK djch m i n g phdi vdi trieu chflng sd't.
3. Khdng cd md'i lien quan gifla PCR - BK dich mang phdi vdi tinh c h i t khdi phat cua benh.
4. Khdng cd md'i lien quan gifla PCR - BK dich mang phdi vdi phan flng Mantoux.
5. Khdng cd md'i lien quan gifla PCR - BK djch mang phdi vdi td'c dp lang m l u .
TAI LIEU THAM KHAO
1. Le Thanh Chfldng (2004). Nhan xet dac diem l l m sang va can l l m sing cua benh n h i n TDMP do lao dieu trj tai khoa Hd h i p benh vien Bach Mai tfl t h i n g 1/2002 - thang 12/2003. Luan van td't nghiep b i c sT Y khoa, tn/dng Dai hpc Y Ha Npi.
2. D6 Chau Hiing ( 1995). Gdp phan nghien cflu d c dac diem lam sang, XQ v l bien ddi mdt sd chi tieu sinh hda, te b i o trong TDMP thanh td do lao. Luan van td't nghiep Thae sT khoa hpc Y Dfldc.
3. Dang Hung M i n h (2002). Hieu q u i cfla sinh t h i i t mang phdi bang kim Castelain dfldi dinh vi ctia sieu am trong chan doan nguyen nhan TDMP.
Luan van Thae sy Y khoa, Dai hpc Y H I Ndi.
4. Hoang Thi Phu'dng (1999). Nghien cflu hieu qua c h i n doan TDMP thanh td h u y i t do lao bang p h l n flng chudi Polymerase. Luan van Thae sT Y hpc, Dai hpc Y H I Ndi.
5. Au Thanh Tiing (1998). Vai trd thfl nghiem PCR - BK trong c h i n doan lao mang phdi. Luan
TCNCYH Phu trWdng 62 (3) - 2009
van td't nghiep bac sT Ndi tru Dai hpc Y Difpc sang va can lam sang d 23 benh n h i n t r i n djch Thanh phd' Ho chf M i n h . tr. 29 - 34. m i n g phdi do lao v l ung thfl. Hpi t h i o khoa hpc
6. Nguyen Xuan Trieu ( 1994). Dac diem lam benh phdi. Vien lao v l benh phdi, t | p 4, tr. 62.
Summary
D I A G N O S T I C V A L U E OF T H E PCR - BK I N T U B E R C U L O U S PLEURAL E F F U S I O N
Objective: (1) To define the diagnostic value of the PCR - BK for tuberculous pleural effusion. (2) The relation between PCR - BK of pleural fluid and somes clinical and paraclinical features of the tuberculous pleural effusion. Material: 127 patients suffer from tuberculous pleural effusion were treated in the Respiration department of Bachmai hospital. Method: prospective. Results: 32.3% cases have positive PCR - BK. There is not any relation between PCR - BK and somes the clinical, paraclinical features of the
•tuberculous pleural effusion (onset, fever and Mantoux, ESR).
Keyword: tuberculous pleural effusion, polymerase chain reaction - Bacillus de Kok (PCR - BK)
NGHIEN CLfu DAC DIEM LAM S A N G , CAN L A M S A N G VA
VI KHUAN TRONG HEN PHE QUAN BOI N H I § M
Dang Quang H u y \ Tran Hoang Thanh^
•^ ,, . ^Sinh vien Dal hgc Y Ha Ngi -Dai hgc Y Hi Ngi
Muc tieu: (1) Tim hieu dac diem lam sing vi can lim sing eiia HPQ bdi nhiSm. (2) Xic djnh cic loai vi khuan trong HPQ bdi nhidm. Boi tugng va phuang pbap ngbien cdu: hoi cdu 124 benh nhan HT'Q bdi nhiim, trong dd cd 79,8%o nd vi 61,3%o trudng hgp dugc dieu trj tai khoa Hd ha'p benh vien Bach Mai. Ket qua va ket luan: khd thd (87,1 %o) li ly do via vien chinh ciia benh nhin va 13%o trUdng hgp vao vien trong tlnh trang suy hd hi'p dp. HPQ bdi nhidm xay ra quanh nam, khdng ed sU khic biet ve miia. 97,6%o trudng hgp cd tien sd HPQ. 62,9%o trudng hgp cd sd dung thudc dt vi ngda can tai nha (sd dung thud'c dang xjt lal thtfdng xay ra HPQ bdi nhidm the vda vi nang). Trong HPQ bdi nhiim cic tneu chdng thudng gap lin lugt:
khd thd (100%o), ho khac ddm due (76,6%o), 41,1%o cd sdi cd cd (25,8%o), ndi ngat quang (28,2%o). Ran rit li trieu chdng thudng gap nhii khi khim phdl vdi 98,5%o, Hep den la ran ngiy vdi 89,5%o, ran am, ran nd vi n rio phe nang giim chl gap vdi 37,9%o vi 32,3%o tuang dng Ve can lam sing dai da sdbenh nhin deu cd sd lugng bach ciu trung Hnh vi td'c do ling miu tang. 13%o benh nhan suy hd hap cap cd tang PaCO, miu.
23,3%o trudng hgp cay vl khuan duang Hnh d ddm vi 71 % phit hien dugc d lin dy diu tien. Cic vi khuan thudng gap nhat trong HPQ bdi nhiim li: K. pneumonia, A. Baumannilyi P. aeruginosa.
Td khda: ben phe quan boi nhiim
LDATVANDE day nfldc ta la mdt trong nhflng nifdc d Chau A cd Hien nay tren the gidi cd k h o i n g 300 trieu ty le hen cao n h i t . Cung nhfl benh phdi t i c ngfldi m l c hen p h i q u i n va theo thdng ke g i n nghen man tfnh, hen p h i q u i n la ganh nang cua