检验检疫学刊
檢驗檢疫學刊
검험검역학간
INSPECTION AND QUARANTINE SCIENCE
2013年
6期
39-43
,共5页
袁慕云%卓锦雪%许龙岩%陈碧玲%张旺
袁慕雲%卓錦雪%許龍巖%陳碧玲%張旺
원모운%탁금설%허룡암%진벽령%장왕
金黄色葡萄球菌%PCR%焦磷酸测序%耐药%检测
金黃色葡萄毬菌%PCR%焦燐痠測序%耐藥%檢測
금황색포도구균%PCR%초린산측서%내약%검측
Staphylococcus Aureus%PCR%Pyrosequencing%Drug Resistance%Detection
[目的]探讨用norA基因特异检测金黄色葡萄球菌方法及耐药情况。[方法]根据金黄色葡萄球菌的norA基因设计扩增引物和测序引物，先用PCR特异性地扩增目的片段，再制备单链模板在测序引物引导下进行焦磷酸测序，将测序结果与GenBank中的norA基因序列比对进行鉴定；用Vitek2进行药敏试验，分析金黄色葡萄球菌耐药情况。[结果]扩增引物和测序引物表现出良好的特异性，28株金黄色葡萄球菌均扩增出大小113bp的DNA片段，焦磷酸测序结果与norA基因序列100%匹配，而阴性对照菌株均未扩增出DNA条带，焦磷酸测序结果阴性；28株金黄色葡萄球菌有14种耐药谱，青霉素、红霉素、苯唑西林和四环素的耐药率分别为71.4%、35.7%、32.1%和28.6%；6株菌株发生C-T突变，对喹诺酮类抗生素菌敏感。[结论]用norA基因可特异性的检测金黄色葡萄球菌；norA基因第354和377碱基位点的突变和喹诺酮类抗生素耐药性无关。
[目的]探討用norA基因特異檢測金黃色葡萄毬菌方法及耐藥情況。[方法]根據金黃色葡萄毬菌的norA基因設計擴增引物和測序引物，先用PCR特異性地擴增目的片段，再製備單鏈模闆在測序引物引導下進行焦燐痠測序，將測序結果與GenBank中的norA基因序列比對進行鑒定；用Vitek2進行藥敏試驗，分析金黃色葡萄毬菌耐藥情況。[結果]擴增引物和測序引物錶現齣良好的特異性，28株金黃色葡萄毬菌均擴增齣大小113bp的DNA片段，焦燐痠測序結果與norA基因序列100%匹配，而陰性對照菌株均未擴增齣DNA條帶，焦燐痠測序結果陰性；28株金黃色葡萄毬菌有14種耐藥譜，青黴素、紅黴素、苯唑西林和四環素的耐藥率分彆為71.4%、35.7%、32.1%和28.6%；6株菌株髮生C-T突變，對喹諾酮類抗生素菌敏感。[結論]用norA基因可特異性的檢測金黃色葡萄毬菌；norA基因第354和377堿基位點的突變和喹諾酮類抗生素耐藥性無關。
[목적]탐토용norA기인특이검측금황색포도구균방법급내약정황。[방법]근거금황색포도구균적norA기인설계확증인물화측서인물，선용PCR특이성지확증목적편단，재제비단련모판재측서인물인도하진행초린산측서，장측서결과여GenBank중적norA기인서렬비대진행감정；용Vitek2진행약민시험，분석금황색포도구균내약정황。[결과]확증인물화측서인물표현출량호적특이성，28주금황색포도구균균확증출대소113bp적DNA편단，초린산측서결과여norA기인서렬100%필배，이음성대조균주균미확증출DNA조대，초린산측서결과음성；28주금황색포도구균유14충내약보，청매소、홍매소、분서서림화사배소적내약솔분별위71.4%、35.7%、32.1%화28.6%；6주균주발생C-T돌변，대규낙동류항생소균민감。[결론]용norA기인가특이성적검측금황색포도구균；norA기인제354화377감기위점적돌변화규낙동류항생소내약성무관。
The method of inspecting staphylococcus aureus with norA gene was discussed. To design the amplifying probe and sequencing probe according to norA gene were designed. Firstly, the gene fragment was amplified. Then the single strain probe was prepared and the pyrophosphoric acid sequencing, implemented and the seguence, compare with the one in GeneBank. Carry out the medicine sensitivity was tested and the drug resistance, analyzed Amplifying and sequencing probes both showed good specificity, and all of the 28 strains got a DNA fragment of 113bp. Pyrophosphoric acid sequencing matched the norA gene sequence with a degree of 100 percent. The negative controls showed the opposite. The 28 strains showed 14 types of drug resistance. The resistance rates of penicillin, erythrocin, oxacillin and tetracycline were 71.4%, 35.7%, 32.1% and 28.6% respectively. Six strains had C-T mutations, and were sensitive to quinolones. The norA gene could be used to inspect staphylococcus aureus. The 354 and 377 base mutations had little relation with resistance to quinolones.
