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目的:研究 ESBLs表型与基因型的关系。方法:（1）药敏试验:K-B法,（2）ESBLs表型确认:双纸片协同法,（3）基因型检测:分别以TEM、SHV通用引物设计PCR体系,行PCR扩增,产物进行分段测序,由测序软件作全基因拼接。结果:1、3、4号菌株为Epn,2号菌株为Eco,其中2、3号株来自同一患儿的不同部位。4株均为TEM亚型,2、3、4号株之序列与国内已报道的TEM序列完全一致,1号株测得序列与2、3、4号株TEM序列有一个碱基因的差别。讨论:苏州地区至少存在两种TEM基因型。4株菌表型耐药状况严重,临床治疗效果不乐观。2、3号株来源于同一患儿不同部位,虽不同菌种,但耐药质粒为同一基因亚型,不排除不同种菌株之间的质粒传播的可能性。PCR及其产物的测序为ESBLs的研究提供了一种有效手段。
Objective: To study the relationship between ESBLs phenotypes and genotypes. Methods: (1) Susceptibility test: KB method, (2) ESBLs phenotype confirmation: double paper synergy method, (3) genotype detection: PCR system was designed by universal primers of TEM and SHV respectively. Segment sequencing, sequencing software for whole genome splicing. Results: The strains Nos. 1, 3 and 4 were Epn and the strains No. 2 were Eco. The strains Nos. 2 and 3 originated from different parts of the same child. 4 strains were all TEM subtype. The sequences of strains 2, 3 and 4 were completely identical with the reported TEM sequences in China. The difference between the sequences of strains 1 and 2 and the TEM sequences of strains 2, 3 and 4 was one base. Discussion: There are at least two TEM genotypes in Suzhou. 4 strains of phenotypic drug resistance is serious, clinical treatment is not optimistic. 2,3 strains derived from different parts of the same child, although different strains, but the same plasmid is the same gene, does not rule out the possibility of plasmid spread between different strains. PCR and sequencing of its products provide an effective method for the research of ESBLs.