摘要
目的建立采用以SYBR GreenⅠ为荧光染料的RT-PCR定量检测MRSA中mecA基因表达的方法,并探讨mecA基因表达水平与甲氧西林耐药水平的关系。方法以RT-PCR同时检测59株临床分离的MRSA中mecA基因和16S rRNA基因的表达量,然后根据公式计算出每株菌mecA基因的相对表达量F值,最后采用SPSS软件以每株菌的苯唑西林MIC值和F值做箱图来观察mecA基因表达水平与甲氧西林耐药水平的关系。结果 RT-PCR定量检测mecA基因和16S rRNA基因的标准曲线方程分别为Y=-3.62X+27.305和Y=-3.54X+27.726,mecA基因的相对表达量F值并没有随着苯唑西林MIC升高而增加。结论 RT-PCR结合SYBR GreenⅠ荧光染料定量检测mecA基因表达是一种相对低廉且可行的方法,mecA基因的表达水平与葡萄球菌属对甲氧西林的耐药水平并无明显的相关性。
OBJECTIVE To establish a method of quantitation of mecA transcription in methicillin-resistant Staphylococcus aureus by RT-PCR which used fluorescent dye SYBR Green Ⅰ and investigate relationship between mecA,transcription and the level of methicillin resistance.METHODS mecA transcription and 16S rRNA transcription were quantified by RT-PCR in 59 strains of methicillin-resistant S.aureus clinical isolates,then the relative transcription of F value was calculated by formula,eventually investigate relationship between mecA transcription and the level of methicillin resistance by Boxplot.RESULTS The standard curvilinear equation of mecA and 16S rRNA which quantitied by RT-PCR were Y=-3.62 X＋27.305 and Y=-3.54X＋27.726 respectively,the relative transcription of F value did not increase when methicillin MIC increased.CONCLUSION Fluorescent dye SYBR Green Ⅰ can be used in quantitation of mecA by RT-PCR.The level of methicillin resistance has no relation with mecA transcription.
出处
《中华医院感染学杂志》
CAS
CSCD
北大核心
2011年第8期1511-1513,共3页
Chinese Journal of Nosocomiology
关键词
金黄色葡萄球菌
逆转录
甲氧西林
抗药性
聚合酶链反应
Staphylococcus aureus
Reverse transcription
Methicillin
Drug resistance
Polymerase chain reaction
