PHAN B 6 CAC GEN MA HOA COAGULASE 6^ CAC GENOTYPE
CLIA STAPHYLOCOCCUS AUREUS
Le Khdnh Trdm\ Dlnh HIPU Dung^
Phgm Thj Thiiy Duang*, Nguyin VQ Trung*
^Bdnh vidn Bach Mai. 'Tmdng Dpi hpc Y Hd Npi Staphylococcus aureus Id cdn nguydn cua nhiiu lopi nhiim trung khde nhau, tmng dd cd nhung bdnh rit npng. Xdc djnh cdc gen quy dinh yiu td ddc luc ed y nghia quan trpng tmng chin dodn, nghien cuv vi ca chi gay bpnh, djch ti hpc phdn tw. Nghidn cdu nhim xdc djnh swphdn bd cde gen md hda eoagulase d cdc genotype ciia S. aureus phdn ldp tw nhdn vidn Idm vide tai ea sd dich vu dn udng d Hd Ndi- Kit qud cho thiy sdu genotype cua gen ma hda eoagulase dd dwac xdc dfnh. cd 1 genotype chiim wu thi (66.2%). cdc genotype phdn bd vdi ty Id khde nhau d cdc chdng phan ldp dwac tw mui vd tay. cd 91.2% ngwdi mang S.
aureus vdi 1 lopi genotype. 8.8% ngwdi mang S. aureus vdi 2 hodc 3 loai genotype. Sdu genotype cua gen ma hda eoagulase da dwac xdc djnh. genotype 6 chiim wu thi (66.2%). Cd 91.2% ngwdi mang S. aureus vdi 1 lopi genotype. Mdt ngwdi cd thi mang S. aureus vdn han 1 lopi genotype.
Tip khda: S. aureus, genotype, gen md hda eoagulase
I. DAT
V A N D ^Staphylococcus aureus la mOt trong nh&ng vi khuln hay gdp nhit tren ca t h l ngudi.
S.aureus cung la can nguyen eiia nhilu loai nhilm triing khae nhau, trong dd thudng gap la nhiem trung ngodi da, ngO dOc Uigc phim, viem xuang tuy, nhilm khuln huylt, nhiem trimg bOnh viOn [2]. Vj tri cu tru hay gdp nhit ciia S.aureus la Id mui trudc. S.aureus ed t h l cu trii thudng xuyen. khdng thudng xuyOn va khOng cu trii trOn ea t h l ngudi [1]. Coagulase Id mOt trong nh&ng y l u td dOc lgc ngogi bao ciia S.aureus. Day Id enzyme lam dOng huylt tuang vd la mOt chl dilm d l phdn biet S. aureus vdi eae tu c l u gdy benh khae [9].
Hien nay. eac nghien cuu v l dieh te hoc phan tu ciia S. aureus cdn rat han c h l . Gen ma hda eoagulase cd tinh da hinh thdi. Viee phdn loai dga vao gen md hda coagulase dan gian, cd do nhay vd do dge hieu eao [5]. VI vdy, chimg tdi filn hdnh d l tai nghiOn c&u nay vdi muc tieu: Xdc djnh sg phan bd eac gen md hda eoagulase d ede genotype eiia S.aureus phdn
ldp tu nhdn viOn lam vlOe tai ca sd djch vu dn udng d Ha NOI.
II. 0 6 l TU'aNG VA PHU'QNG PHAP
1. Ddi tugng
47 chiing S. aureus phdn ldp dugc tu tay vd mui cua 103 nhdn viOn ldm viOc tgi 5 ca sd djch vu dn udng d Ha NOi, tilp xiic trge filp vdi th&c dn. Thdi gian nghien c&u: t& thdng 3/2009 d i n .9/2010.
2. Phuang phap
Lly benh phim bing tdm bOng vO triing d ea 2 bdn tay vd 2 Id mui ngodi 4 lln, cdch nhau 1 tuin. Nudi c l y phdn ldp, luu gi& chiing [1]. Tdch DNA. Tiln hdnh ky thudt PCR [6], elt bdng enzyme Alul (hdng Fermentas, Canada), mdy PCR (Mastercycler gradient, Eppdorf, D&c), hO thdng diOn di (Bp diOn di ngang Horizon®58-Gibeo-BRL, Anh), hO thdng chgp anh (mdy Geldoe, hang BioRad, Phdp).
4 logi mdi:
C0AG1:5'-ATACTCAACCGACGACACCG-3'
C0AG2: 5'-GATTTTGGATGAAGCGGATT-3' C0AG3: 5'-CGAGACCAAGATTCAACAAG-3' C0AG4: 5'-AAAGAAAACCACTCACATCA-3'
TiOu chuln x l p genotype: ede genotype eua S. aureus dugc xdc djnh dga vdo: kich ed eua bdng DNA duge elt bdng enzyme Alul, sd lugng ede dogn DNA sau khi elt. Genotype 1:
465bp; genotype 2: 465bp, 800bp; genotype 3: 465bp, 250bp; genotype 4: 465bp, 550bp;
genotype 5: 465bp, 85bp ; genotype 6: 465bp, 250bp, 165bp,
- Cdc sd liOu dd dugc xu ly b i n g p h i n Bdng 1. Phdn bd cdc genotype cua gen
m i m SPSS 16.0.
III. K^T QUA
1. Phdn bd cdc genotype ciia gen ma hda coagulase
Trong sd 47 ehiing S.aureus phdn ldp dugc, cd 6 genotype eiia gen md hda cho coagulase. Trong dd, genotype 2, 3, 4 vd 5 cd 2 s i n phim elt. Nhdm 6 ed 3 san phim e l t Cd 31 ehiing (66,2%) thudc genotype 6. Tit ca cdc genotype d i u ed san phim kieh cd 465bp.
md hda coagulase d 47 chiing S.aureus Geno
-type 1 2 3 4 5 6
Sdsp 1 2 2 2 2 3
KIch c d 800
+
sdn p h i m sau cdt bdng Alul (bp) 550 465 250 165
+ +
TT ' ^
+ + +
+ + + 85
+
n (%) 5(10.6) 6(12,7) 3 (6,3) 1 (2.1) 1 (2.1) 31(66.2) Bang 2. Phdn bd cac genotype gen ma hda coagulase ciia S. aureus a tay va mui
Genotype
Vj tri cdc Chung S.aureus dugc phdn ldp
Tdng sd
1 2 3 4 5 6 Cdng Kit qua d 9/14 Chung's.,
Tay 1 3 0 1 0 9 14
bang 2 eho thiy, genotype 6 cd mdt d aureus d tay. Tilp theo la cde genotype
Mui 4 3 3 0 1 22 33
22/33 ehiing S.aureus 1, 2, 3, 4 va 5 vdi edety
5 6 3 1 1 31 47
phdn lap tu mui vd le khde nhau.
TCNCYH 78 (1) - 2012
2. Phan bd sd lugng cdc genotype ma hda coagulase
4 9 11 14 15 18 23 25 M 27 29 32 33 35 36 37 41
465 bp 250 bp
165 bp
Hinh 1. Ket qua cat ADN cua -not so chiing S. aureus bdng enzyme Alul M: Marker
Cae ehung edn lai Id S. aureus thuOc cdc genotype khde nhau
K i t qua d hlnh 1 eho thay, phdn Idn cdc chiing c6 3 san pham elt ed kich cd 465, 250 vd 165bp (thuOc genotype 6).
. Bang 3. Phan bd nhan vien theo sd lugng cdc genotype gen ma hda coagulase ciia S.aureus
Sd lugng genotype 1
2 3 4 COng
Sd nhan vien"
21 1 1 0 23
(%) 91,2
4,4 4,4 0,0 100,0 Ghi chu:' Chi tinh sw cd mat cua S. aureus d nhin vidn khdng tinh din lin liy miu.
Trong sd 23 nhan vien, sd ngudi mang S. aureus vdi 1 genotype eua gen ma hda eoagulase hay gap nhit vdi ty le 91,2%.
IV BAN LUAN
Phan bd cdc genotype cua gen md hda coagulase: Viee xae djnh ty 10 mang S.aureus la ea sd d l danh gia djch t l hoc phdn tu cdc Chung S.aureus. Theo k i t qua d bang 1,
chiing tOi dd xdc djnh dugc 6 genotype eiia gen md hda eoagulase dga vdo ede san pham elt bdng enzym Alul. Trong nghiOn c&u ndy.
cd t i t ea 6 dogn e l t ed kieh ed lan lugt Id 85.
165. 250, 465, 550 vd 800bp duge xdc djnh.
Sd cdc dogn elt tu 1 d i n 3, trong dd cd 4 kilu
cdch ed 2 dogn elt. 0$c biOt, genotype 6 gdp phd biln nhit vdi 3 dogn elt, chilm ty 10 66,2% tdng sd chung S.aureus phdn ldp duge. Theo nghiOn c&u eua Schwarzkopf vd cOng sg trOn 30 ehiing S.aureus phdn ldp tu bOnh phim ldm sdng, mOi ciia bOnh nhdn vd nhdn viOn y t l , cung ed 6 genotype dugc phdt hiOn. Sg da dgng ndy dugc xdc djnh khOng nh&ng d cdc chung S.aureus phdn ldp tu ngudi md ea nh&ng chung phdn ldp tu dOng vdt, vd tgi nh&ng vung dja ly khde nhau [3,7], Trong nghiOn c&u eiia ehiing tOi. genotype 6 chilm ty 10 66.2%. Oilu ndy eho thiy cd sg chiem uu t h l cua mOt ehiing S.aureus mang genotype 6.
Bang 2 cho thiy cde genotype phdn bd vdi ty le khde nhau d cdc chiing phdn ldp duge tu mui vd tay, Tuy nhiOn vl sd lugng mdi loai genotype eiia ede chiing phdn ldp dugc chua dii Idn d l xae djnh y nghTa thdng ke cua sg khac blot nay,
Phan bd sd lugng cdc genotype ma hda coagulase: Nlu chi tinh sg cd mdt cua cdc chung vd genotype cua nd trOn 23 nhdn vien, kit qua d bang 3 cho thiy, ed 91.2% ed 1 genotype ciia cdc chung phdn ldp dugc sau 4 lan lly miu. Cd 1 trudng hgp mang 2 genotype vd 1 trudng hgp mang 3 genotype trOn cimg 1 nhdn viOn, Khi phdn tich v l genotype ciia 4 ehiing ndy, ehiing ed tdi 3 kilu gen khde nhau, Oilu ndy cung cd t h l dugc giai thich do cdc chiing S. aureus khde nhau cung cu trii. MOt sd nghiOn c&u trudc ddy cQng cho thiy, d nh&ng ngudi md S.
aureus cu trii theo kilu cdch thudng xuyOn thl thudng ed 1 chiing xult hiOn. Kilu cdch cu trii khOng thudng xuyOn hay do nhilu chiing S.
aureus trOn mOt ed t h l tgo ra [3]. Cde chiing hay eae genotype eiia S. aureus chilm uu i n l vd trd thdnh cde ehiing tgo ra kilu cdch cu trii thudng xuyen thudng Id cde ehiing phdt triln
mgnh, ed y l u td dOe Igc eao, k h i ndng thieh nghi Idn vd l l n dt cdc ehiing khde d l phdt triln [6, 9]. Ngudi ta thiy ring, xOt v l gdc dO da dgng ciia kilu gen, viOc xdc djnh cdc genotype cua S. aureus di d u g g chung minh Id mOt phuang fiOn dan giln, ehlnh xdc vd rit ed gid trj trong viOc phdn loai cdc chiing S.aureus [5, 7].
V. K^T LUAN
Sdu genotype eiia gen md hda coagulase eiia S. aureus di dwgc xic djnh, genotype 6 chilm uu t h l (66.2%).
91.2% ngudi mang vi khuln cd 1 logi genotype. MOt ngudi ed t h l mang S. aureus vdi han 1 logi genotype.
TAI
LIEU THAM
K H A O1. Le Huy Chinh (2009) Tg c l u vdng. Vi khuan y hoe. Nha xult ban Giao due ViOt Nam, 45- 56.
2. Bill Khde Hdu va cgng sg (2002) NghiOn cuu tinh khdng thude khdng sinh tu S.
aureus phdn ldp t& v l t md nhilm triing. Tgp chi Y hpc thgc hdnh, sd 9, 54-56.
3. Aarestrup, P.M., C A . Dangler, and L.M. Sordillo (1995). Prevalence of eoagulase gene polymorphism in Staphylococcus aureus isolates causing bovine mastifis. Can J Vet Res,59(2): 124-128.
4. Deresinski, S (2009). Vancomycin in combination with other anfibiofics for the treatment of serious methieillin-resistant Staphylococcus aureus infections. Clin Infect Dis, 49(7): 1072-1079.
5. Goh, S.H., et al (1992). Molecular typing of Staphylococcus aureus on the basis of eoagulase gene polymorphisms. J Clin Microbiol, 30(7): 1642-512.
6. Leman, R, et ai (2004). Nasal carriage of methieillin-resistant Staphylococcus aureus
TCNCYH 78 (1) - 2012
in an American Indian populafion. Infect Control Hosp Epidemiol, 25 (2), 121-518.
7. Schwarzkopf, A. and H. Karch (1994).
Genetic variafion in Staphylococcus aureus coagulase genes: potenfial and limits for use as epidemiological-marker. J Clin Microbiol, 32(10): 2407-2412.
8. Van Belkum, A., et al (2009).
Reclassificafion of Staphylococcus aureus nasal carriage types. J Infect Dis, 199 (12):
1820-1826,
9. Vanden Bergh, M.F., et al (1999).
Follow-up of Staphylococcus aureus nasal carriage after 8 years: redefining the persistent carrier state, J Clin Microbiol, 37 (10): 3133-3140.
Summary
DISTRIBUTION OF COAGULASE GENE IN GENOTYPES OF STAPHYLOCOCCUS AUREUS
Staphylococcus aureus is a cause of many different infections, including life-threatening infecfions. Detection of virulent genes plays an important role in diagnosis, study on pathogenesis and molecular epidemiology. The study's objective was to recognize the distribufion of coagulase gene in genotypes of Staphylococcus aureus strains isolated from noses and hands of staffs working in restaurants in Hanoi. Results showed that 6 genotypes of coagulase gene were determined with one genotype predominafing the other five (66.2%). There were different distribufions of eoagulase gene in S. aureus isolated from noses and hands, 91.2% of S. aureus- harboring staffs had one genotype. 8,8% of S, aureus-harboring staffs had 2 or 3 genotypes. In conclusion, 6 genotypes of eoagulase gene were determined with genotype 6 predominafing the other five (66.2%). 91.2% of S.aureus-harboring staffs had one genotype. There were possibilities of having more than one S. aureus genotype In a person.
Keywords: S. aureus, genotype,^ene coding coagulase
DOT BI^N GEN COL1A1 O BENH NHI TAO XL^aNG B A T TOAN
Hd e l m T u \ Trln Van Khdnii\ Biii Thj Hdng Chau\
Trln Huy Thjnh\ Le Hodi Chuang^, Ta Thanh Van^
^Trwdng Dpi hpc Y Hd Npi, 'Bdnh vidn Php sdn Tmng wang Bpnh tpo xuxyng bit todn Id bpnh di truyin trdi tren nhiem sac thi thwdng. Phin Idn cdc ddt biin Idm
thay ddi bd ba acid amin Gly-X-Y, pha vd ciu true cua pmtein Collagene I. Nghien cwu dwac thuc hipn nham phdt hipn cdc dang dpt biin gen G0L1A1 tren bpnh nhi mac bpnh tpo xwang bit todn. Kit qua nghidn cwu cho thiy 11/15 bdnh nhi dwac phdt hipn cd ddt biin dim trdn gen C0L1A1. Kit lupn: Da phdt hipn thdnh edng dot biin dim trdn bpnh nhi Tpo xwang bat todn Vidt Nam vdi ty lp 11/15 bpnh nhi.
TIP khoa: benh tao xuang bit toan - 01, dpt biln gen C0L1 Al
