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目的 建立快速检测非培养感染物中抗生素耐药基因表达的分子生物学方法 ,为临床的早期诊断和治疗及研究微生物学耐药机制提供一种理想的分子生物学技术。方法 应用聚合酶链反应测定 7种抗生素耐药基因 ,并通过优化测定体系中镁离子浓度、引物浓度及引物退火温度等 ,对PCR反应方法进行了质控。结果 7种抗生素耐药基因扩增条带清晰 ,无非特异性扩增条带 ,与所设计的扩增片段大小相符。结论 应用PCR检测微生物的耐药基因 ,灵敏度高 ,特异性强。可简便、快速地检测细菌、真菌、衣原体和支原体的耐药基因 ,尤其联合检验对临床早期诊断、治疗及鉴定耐药菌株及亚型具有一定价值。
Objective To establish a molecular biological method for rapidly detecting the expression of antibiotic resistance genes in non-cultured infectious agents, and to provide an ideal molecular biology technique for early clinical diagnosis and treatment and for studying the mechanism of microbial resistance. Methods Seven kinds of antibiotic resistance genes were detected by polymerase chain reaction (PCR). The quality of PCR reaction was controlled by optimizing the concentration of magnesium ion, primer concentration and primer annealing temperature in the system. Results The bands of 7 antibiotic resistance genes were clear and non-specific amplification bands were consistent with the size of the amplified fragments. Conclusion PCR detection of microbial resistance genes, high sensitivity and specificity. Can be easily and quickly detection of bacteria, fungi, chlamydia and mycoplasma resistance genes, especially the joint test early clinical diagnosis, treatment and identification of resistant strains and subtypes has a certain value.