中华检验医学杂志
中華檢驗醫學雜誌
중화검험의학잡지
CHINESE JOURNAL OF LABORATORY MEDICINE
2008年
10期
1119-1123
,共5页
念珠菌,白色%氟康唑%抗药性,真菌%细胞色素P450酶系统%氧化还原酶类%真菌蛋白质类%突变
唸珠菌,白色%氟康唑%抗藥性,真菌%細胞色素P450酶繫統%氧化還原酶類%真菌蛋白質類%突變
념주균,백색%불강서%항약성,진균%세포색소P450매계통%양화환원매류%진균단백질류%돌변
Candida albicans%Fluconazole%Drug resistance,fangal%Cytochrome p-450 enzyme system%Oxidoreductages%Fangal proteins%Mutation基金项目:国家自然科学基金资助项目(30471563)
目的 筛查耐氟康唑白念珠菌ERG11基因突变,探讨其与耐药的关系.方法 用柯玛嘉显色培养基和25S rDNA转座内含子保守区分型方法,收集鉴定白念珠菌临床株.联用微量稀释法和Rosco药敏纸片扩散法体外测定试验菌株对氟康唑的敏感性.以ERG11序列明确的白念珠菌标准耐药株ATCC 76615-19和达令顿株为质控,以标准敏感株C1b和临床敏感株02928为对照,分3段扩增临床耐药株ERG11基因并测序.结果 获得15株A型白念珠菌临床耐药株.ERG11基因测序共发现16处同义突变和11处错义突变.质控株突变与既往报道一致.敏感株共出现9处同义突变和3处错义突变G640A(E165K)、A945C(E266D)、G1609A/G(V488I).耐药临床株中的14株均出现2处共有的错义突变G487T(A114S)和T916C(Y257H),不伴其他突变;另1株出现8处同义突变和4处错义突变R541C(Y132H)、T495A(D116E)、A530C(K128T)、T1493A(F449Y),其中T1493A(F449Y)未见报道.结论不同来源的白念珠菌临床耐药株集中发生G487T(A114S)和T916C(Y257H)突变,暗示这2处突变极可能参与菌株耐药.耐药株可以发生T1493A(F449Y)突变.
目的 篩查耐氟康唑白唸珠菌ERG11基因突變,探討其與耐藥的關繫.方法 用柯瑪嘉顯色培養基和25S rDNA轉座內含子保守區分型方法,收集鑒定白唸珠菌臨床株.聯用微量稀釋法和Rosco藥敏紙片擴散法體外測定試驗菌株對氟康唑的敏感性.以ERG11序列明確的白唸珠菌標準耐藥株ATCC 76615-19和達令頓株為質控,以標準敏感株C1b和臨床敏感株02928為對照,分3段擴增臨床耐藥株ERG11基因併測序.結果 穫得15株A型白唸珠菌臨床耐藥株.ERG11基因測序共髮現16處同義突變和11處錯義突變.質控株突變與既往報道一緻.敏感株共齣現9處同義突變和3處錯義突變G640A(E165K)、A945C(E266D)、G1609A/G(V488I).耐藥臨床株中的14株均齣現2處共有的錯義突變G487T(A114S)和T916C(Y257H),不伴其他突變;另1株齣現8處同義突變和4處錯義突變R541C(Y132H)、T495A(D116E)、A530C(K128T)、T1493A(F449Y),其中T1493A(F449Y)未見報道.結論不同來源的白唸珠菌臨床耐藥株集中髮生G487T(A114S)和T916C(Y257H)突變,暗示這2處突變極可能參與菌株耐藥.耐藥株可以髮生T1493A(F449Y)突變.
목적 사사내불강서백념주균ERG11기인돌변,탐토기여내약적관계.방법 용가마가현색배양기화25S rDNA전좌내함자보수구분형방법,수집감정백념주균림상주.련용미량희석법화Rosco약민지편확산법체외측정시험균주대불강서적민감성.이ERG11서렬명학적백념주균표준내약주ATCC 76615-19화체령돈주위질공,이표준민감주C1b화림상민감주02928위대조,분3단확증림상내약주ERG11기인병측서.결과 획득15주A형백념주균림상내약주.ERG11기인측서공발현16처동의돌변화11처착의돌변.질공주돌변여기왕보도일치.민감주공출현9처동의돌변화3처착의돌변G640A(E165K)、A945C(E266D)、G1609A/G(V488I).내약림상주중적14주균출현2처공유적착의돌변G487T(A114S)화T916C(Y257H),불반기타돌변;령1주출현8처동의돌변화4처착의돌변R541C(Y132H)、T495A(D116E)、A530C(K128T)、T1493A(F449Y),기중T1493A(F449Y)미견보도.결론불동래원적백념주균림상내약주집중발생G487T(A114S)화T916C(Y257H)돌변,암시저2처돌변겁가능삼여균주내약.내약주가이발생T1493A(F449Y)돌변.
Objeetive To detect ERG11 gene mutations in clinical isolares of Candida albicans resistant to fluconazole.and discuss their relationship with formation of drug resistance.Methods Clinical specimens were collected.CHROMagar mediuln and amplification of the fragment spanning the conserved sequence of 25S rDNA including some transposable introns.were used to identify subtype Candida albicans isolates.FIuconazole sensitivity was detected in vitro through microdilution and Rosco tablets.The other three fragment of ERG11 gene were amplified and followed by sequencing with resistant type strain ATCC 76615-19 and Candida albicans Darlington strain with two sensitive isolates as controh.Results Fifteen resistant isolates of Candida albicans were found,all of which were type A.Sixteen silent mutations and 11 missense mutations were detected.Mutations in ATCC 76615-19 and Darlington strain were same with what had been reported.In the 2 sensitive strains.G640A(E165K),A945C(E266D)and G1609A/G(V488I)occurred,as well as the other 9 silent mutations.Only G487T(A114S)and T916C(Y257H)existed in each of 14 resistant isolates.In the other one resistant isolate,T541C(Y132H),T495A(D116E),A530C (K128T)and T1493A(F449Y)occurred Mong with 8 silent mutations.Conclusions The occurrence of G487T(A114S)and 1916C(Y257H)in 14 isolates from different sources suggested they may involve in fluconazole resistance.The novel mutation T1493A(F449Y)can appear in resistant isolves of Candida albicans.
