39 2 Vol.39No.2 20182 ActaEcologiaeAnimalisDomastici Feb.2018 MRSA! "#$%&' MLST 1, 1, 1, 2, 1, 3, 1 (1., 832003;2.!"#$ %&',!" 843000; 3. () *+,-./0,1 *+ 730046) [ (]!"#$%&' () MRSA *+,-./ -.01,23456789:" ; 71, <= -./,>? MRSA @+AB;C3D PCR E8? MRSA *+,@+-.01 9: MLST,71, <= 9F13, MRSA,9FG!18.3% 13, MRSA *+,H9 F 犿犲犮犃 I 犾犻狀犃 -.01,9FG!100.0%;7,9F 狋犲狋犿 -.01,9FG!53.8%;4, 9F 犿狊狉犅 -. 0 1,9 F G! 30.8%;4, 9 F 犃犪犮犃 犇犪 -. 0 1,9 F G! 30.8%; MLST J K,13, MRSA L! 6 M ST, N! ST188 ST9 ST63 ST2700 ST968 ST2373,O ST9 *+!!$%&' MRSA *+, -. P QR"ST0U,!VW XS3.QR"Y [)*+] - <=;-.01;MLST [,-./] S811.5 [01 ] A [0 /] 1005 5228(2018)02 0066 05 234567 89):; & <,=>?@ABCD( 犛狋犪狆犺狔犾狅犮狅犮犮狌狊犪狌狉犲狌狊,SA)E 5 2 3 4 & D < [1] F G >? @ A B C D ( 犕犲 狋犺犻犮犻犾犻狀 狉犲狊犻狊狋犪狀狋犛狋犪狆犺狔犾狅犮狅犮犮狌狊犪狌狉犲狌狊,MRSA) 5<H IJKL M &,NO KL PQRSTUV KLWXY YM Z[\ ]^[2 3] _[,` abc ) V D, SA D Y,Y M M D I J, M IJ [4 6] >?@ABCD D PQ Z H, P 0, = [7 8] _[./:, MRSA RS, Q RS, L./ MRSA b I, MRSA D S D 1998,Maiden [9] YX ( 犿狌犾狋犻犾狅犮狌狊狊犲狇狌犲狀犮犲狋狔狆犻狀犵, MLST), Y =YX (MLST) < CRS&, TU R S&./ [10 11]. / b MRSA RS MLST [ MRSA R S R S &, ' MRSA bic R S, - D M 1 1.1 234 Baird Parker 8 BHI 8!" #$%;&'D ( 犈. 犮狅 犾犻 )DH5αDH ; D DNA [()*] 2016 12 08 [ >+,] -. /01I (2016BC001); M.: 2 (2016YFD0500900); $./ +,(XJGRI2016040) [3456] 7 8(1990-),9,:; +I,./,./ & E mail:2393802294@qq.com [ 34] 1(1971-),<,=F>GI,",?@, AcB&./ E mail:qj710625@163.com
2 7 8, : 234 MRSA RS MLST./ 67 CDE pmd19 T,DNA Marker TaKaRa$%; F + G H E #$%; D IJ F+H E #$%;KL M C D E DNA N O ( E ( ) #$% 1.2 56&78 P QR STU4V#$ WX Y 2 3 4 2 221, Z[\]23^ 75%_`aC 2b Sc,( 3 P D E, P, S 8 1.3 9:; 2 H P Baird Parker 8, P37 Z 812~48h D D,b 8 8 S 8 D S J * L@ ` D SA Q, GenBank $ >? @ A B C D 16 犛狉 RNA, SA S16 犛狉 RNA, >?@ABC D >?@ABCDb BHI 37 824h, 500μL 0 D 500 μl40%1 20% 1 D,- 20 1.4 # 2 MRSA D H D VD, HP BHI 8 P37 820~24h, D [ D 0.5,D 200μL P MHA 8, 14 H I J, I J Q 8 37 Z 8 8 48h D < PCR 犿犲犮犃 [ MRSA D 30μgb F D 21mm D MRSA 1.5 "#$%&' YM PCR J ( 犿狊狉犃 犿狊狉犅 ) J ( 犲狉犿犃 犲狉犿犅 犲狉犿犆 ) ( 狏犪狋犃 狏犪狋犅 狏犪狋犆 ) J 犪犪犮犃 犇 J ( 狋犲狋犽 狋犲狋犿 ) G J 犾犻狀犃 S 1.6!$%< DNA => D H D V D H D BHI 8 P37,D1.5mL D 8 P D EP b 12000r/min, \, 3 D DNA C D E 3 S DNA C D,4μLPCR 1% NO,EB L@ bno DNA X P -20, P MLST./ 1.7?@ABC (MLST) MLST. / 犪狉犮犆 犪狉狅犈 犵犾狆犉 犵犿犽 狆狋犪 狋狆犻 狔狇犻犾 7 V, S PCR, S, < V, K S!, " S ^#,C$% & (htp://www.mlst.net)' D VX X,^ D7V X ' (sequencetyp ing,sts) 2 2.1 DE 9:; Baird Parker 8 8, 221 ABC D 71 SA,b Baird Parker 8 @ D 1 < 16 犛狉 RNA PCR 2, 32.1% -1 SA F Baird ParkerGH$& IJ Fig.1 Thecolonymorphologyof SAinBaird Parker -2 16 犛狉犚犖犃 $%& PCR KL Fig.2 Amplificationof16 犛狉犚犖犃 genebypcr M.Marker2000;1 10.Isolates
68 c 3 39 2.2 MRSA &:; # 2 71 SA 13 MRSA,mecA PCR 3, 18.3%,Q P3, 2,QR4,STU4 13 MRSA b F D MP21mm, <,0 MRSA,b F ( )*,+,, 7,2,1 -,MRSA b F ( 1-3 犿犲犮犃 $%& PCR KL M.Mark2000;1 3,6 7,11 13,16 17,20 22:PCR Fig.3 Amplificationof 犿犲犮犃 genebypcr M.DNA MarkerDL 2000;1 3,6 7,11 13,16 17,20 22:PCRproducts M113! MRSA &"#M N Table1 Theanalysisresultofresistancephenotypeinthe13strainsofMRSA Resistancephenotype Resistancephenotype SA xj 06 犘犈犖, 犆犉犡, 犜犕犘 SA xj 27 犘犈犖, 犆犉犡, 犜犕犘, 犈犕 SA xj 08 犘犈犖, 犆犉犡, 犜犕犘, 犈犕 SA xj 41 犘犈犖, 犆犉犡, 犜犕犘, 犜犈犜 SA xj 10 犘犈犖, 犆犉犡, 犜犕犘, 犈犕 SA xj 42 犘犈犖, 犆犉犡, 犜犕犘, 犈犕, 犜犈犜, 犌犈犖 SA xj 13 犘犈犖, 犆犉犡, 犜犕犘, 犈犕 SA xj 43 犘犈犖, 犆犉犡, 犜犕犘, 犈犕 SA xj 14 犘犈犖, 犆犉犡, 犜犕犘 SA xj 59 犘犈犖, 犆犉犡, 犜犕犘 SA xj 21 犘犈犖, 犆犉犡, 犜犕犘 SA xj 61 犘犈犖, 犆犉犡, 犜犕犘 SA xj 22 犘犈犖, 犆犉犡, 犜犕犘.:PEN. ;CFX.b F ;TMP. (;EM. ;TET. ;GEN.- ; Note:PEN.penicilin;CFX.Cefoxitin;CLI.clindamycin;TMP.Trimethoprim;EM.erythromycin;TET.tetracycline; GEN.gentamicin; 2.3 MRSA "#$%' 13 MRSA / 犿犲犮犃 犾犻狀犃,Q 7 / 狋犲狋犿,4 / M2 13! MRSA O "#$% 犃犪犮犃 Da,4 / 犿狊狉 B,1 / 犞犪狋 C, MSSA,MRSA YM 0 2 Table2 Theresultofcarryingresistancegenesof13strainsofMRSA Z Carryingresistancegene Z Carryingresistancegene SA xj 06 犿犲犮犃, 犾犻狀犃, 犃犪犮犃 犇犪, 狋犲狋犿 SA xj 27 犿犲犮犃, 犾犻狀犃. 狋犲狋犿 SA xj 08 犿犲犮犃, 犾犻狀犃, 狋犲狋犿, 犿狊狉犅 SA xj 41 犿犲犮犃, 犾犻狀犃, 狋犲狋犿 SA xj 10 犿犲犮犃, 犾犻狀犃, 犃犪犮犃 犇犪, 犞犪狋犮, 犿狊狉犅 SA xj 42 犿犲犮犃, 犾犻狀犃, 狋犲狋犿 SA xj 13 犿犲犮犃, 犾犻狀犃, 犃犪犮犃 犇犪, 犿狊狉犅 SA xj 43 犿犲犮犃, 犾犻狀犃, 犃犪犮犃 犇犪, 狋犲狋犿, 犿狊狉犅 SA xj 14 犿犲犮犃, 犾犻狀犃 SA xj 59 犿犲犮犃, 犾犻狀犃 SA xj 21 犿犲犮犃, 犾犻狀犃, 狋犲狋犿 SA xj 61 犿犲犮犃, 犾犻狀犃 SA xj 22 犿犲犮犃, 犾犻狀犃 2.4 MLST 3 MRSA / ST188,ST63,ST9, ST2700,ST968,ST2373,6H MLST,Q 2 ST188, 15.38%,3 ST63, 23.08%,3 ST9, 23.08%,2 ST2700, 15.38%,2 ST968, 15.38%,1 ST2373, 7.69% ( 4) MRSA ST 1 0 3 20 60 : MRSA [,>?@ ABCD(SA) 2 [12]./
2 7 8, : 234 MRSA RS MLST./ 69-4 13! MRSA & MLST Fig.4 TheresultsofMLSTgenotypingof13strainsofMRSA 4,MRSA 5 Z 犿犲犮犃 β J,, ZQ6,7 Q6 D [13 14] SA Y H 8, 9:',[ [15] ;D 1~3 H, ^, ;D Z,<5 ; ^= >,Qb<?@ D, 1 +JD A Bb,^ COD E, + JD ). / :,SA F, <H D G G PHHIHH J 3 [16] +,K 0 D LM K D L N +, [O < SA M,OD YM MR SA KL N D./ 234 MRSA, 6H ( 犿犲犮犃 犾犻狀犃 犃犪犮犃 犇犪 狋犲狋犿 犿狊狉犅 犞犪狋犮 ),0 MRSA 犿犲犮犃 犾犻狀犃,: [17]./ FG 3 MRSA PQO [18]./ b F 3 MRSA. / Y% YH,,^ 50 O 0 1 < P O b, RS TU,= P=O, b Y F, O5 P 6 3 TV,1 OW X Y Z, 8 67, = IJ,,b MLST % & SA 2840 H ST,J( 4929 D,= D Z 91 [eburst V3 )*, D ST b 5 V! \ ] ST97 ST5 ST398 ST1 ST9,! \ ] ST97,!\] ST398,b./ MLST ST188 ST63 ST9 ST2700 ST968 ST2373,Q ST9 ST63 RS P^G [19] 3_ H `ab- MRSA,cP ST9, a MRSAST9 IJ & Kehrenberg [20] I [ I 3I ST9 MRSA D, ST9ObI [21] b D ST188 +,./ ST9 ST188 234 MRSA D b<, I D 1 a<,= D IJ *G a, +, MRSA b [O ba I Bb, + ; MRSA RS./ M 01: [1] BELKUM AV,MELLESDC,NOUWENJ,etal.Co evolu tionaryaspectsofhumancolonizationandinfectionbystaphy lococcusaureus[j].infectgenetevol,2009,9(1):32 47. [2] SOMMERH?USERJ,KLOPPERTB,WOLTER W,etal. Theepidemiology ofstaphylococcusaureusinfectionsfrom subclinicalmastitisindairycowsduringacontrolprogramme [J].Veterinarymicrobiology,2003,96(1):91 102. [3] MAGIORAKOSAP,SRINIVASAN A,CAREYRB,etal. Multidrug resistant,extensively drug resistantand pandrug resistantbacteria:aninternationalexpertproposalforinterim standarddefinitionsforacquiredresistance[j].clin Microbiol Infect,2012,18(3):268 281. [4],,,. # $ 2 3 4 >? @ A B CD [J]. - ;,2011,32(10):59 62. [5], 1,,. >? @ A B C D [J]. L & 3,2011,19 (1):31 38. [6] ZHANG W,HAOZ,WANG Y,etal.Molecularcharacter izationof methicilin resistantstaphylococcusaureusstrains frompetanimalsandveterinarystafinchina[j].theveteri naryjournal,2011,190(2):125 129. [7] MISTRY H,SHARMAP,MAHATOS,etal.Prevalenceand CharacterizationofOxacilinSusceptible meca PositiveClini calisolatesofstaphylococcusaureuscausingbovine Mastitis inindia[j].plosone,2016,11(9):e0162256 [8] TURUTOGLU H,HASOKSUZ M,OZTURK D,etal.Me thicilinandaminoglycosideresistancein Staphylococcusau reusisolatesfrom bovine mastitisandsequenceanalysisof
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