呼吸道感染流感嗜血杆菌生物学分型及耐药基因研究

Study on the biological typing and drug resistance genes of Haemophilus influenzae

目的调查Hi的阳性率、生物学分型和耐药情况。方法 2015年1月~2016年12月从惠东县人民医院门诊及住院患者呼吸道标本中分离流感嗜血杆菌共200株,用法国梅里埃系统API NH进行鉴定确认生物型,并同时检测β-内酰胺酶。采用K-B纸片扩散法对12种抗生素进行药敏实验。根据药敏结果,对于有对β-内酰胺酶类抗生素产生耐药性的Ⅰ、Ⅱ、Ⅲ、Ⅳ型别,PCR检测TEM-1和ROB-1基因。统计不同生物型的β-内酰胺酶、抗生素耐药率、基因检出率。结果共检出Ⅰ~Ⅴ型、Ⅷ型6种生物型,其中比例最高的是Ⅲ型,占38%;其次是Ⅱ型,占25%。Ⅴ型、Ⅷ型未见对12种抗生素耐药。Ⅳ型对于抗生素的耐药率最高,对于氨苄西林、头孢呋辛、头孢他定、氨曲南、阿奇霉素均产生耐药。35.16%的菌株对氨苄西林等β-内酰胺类抗生素呈现耐药性。34.04%菌株产生β-内酰胺酶,TEM-1阳性率为54.26%,ROB-1为6.38%。结论惠东县人民医院收集的呼吸道感染流感嗜血杆菌的生物型有Ⅰ~Ⅴ型、Ⅷ型6种,以Ⅱ型、Ⅲ型为主,Ⅳ型亦占一定比例。Ⅴ型、Ⅷ型未见对12种抗生素耐药。Ⅳ型对于抗生素的耐药率最高。对氨苄西林等β-内酰胺类抗生素,耐药机制以质粒介导的TEM产β-内酰胺酶型为主,少数为ROB-1介导。关注Hi致病生物型的流行情况,对于合理选择抗生素具有重要意义。

Objective To investigate the positive rate,the biological type and the drug resistance of Haemophilus influenzae.Methods 200 strains of Haemophilus influenzae were separated from outpatient and hospitalized patients of Huidong County People′ s Hospital from January 2015 to December 2016.Biological types and beta lactamase were identified with API NH of France bio Merieux identification system.The drug sensitivity test of 12 antibiotics was performed by K-B disk diffusion method.According to the results of drug sensitivity,TEM-1 and ROB-1 gene of the strains of Ⅰ, Ⅱ, Ⅲ,and Ⅳ biotypes were detected by PCR.Results There were 6 biotypes,which were type Ⅰ-Ⅴ,andⅧ type.The highest proportion was type Ⅲ（38%） and type Ⅱ（25%）.Type Ⅴ,type Ⅷ was sensitive to 12 antibiotics.TypeⅣ had the highest antibiotic resistance rate of Ampicillin,Cefuroxime,Ceftazidime,Aztreonam and Azithromycin.35.16%of the strains showed resistance to Ampicillin and other beta lactam antibiotics.34.04% strains produced beta lactam.The positive rate of TEM-1 gene was 54.26%,while the positive rate of ROB-1 gene was 6.38%.Conclusion There were 6biological type of Haemophilus influenzae in Huidong County People′ s Hospital.Type Ⅱ and type Ⅲ were the dominant biotypes,and Ⅳ type also shared a certain proportion.Type Ⅴ,type Ⅷ was sensitive to 12 antibiotics.The biological type which was highest resistance to antibiotics would be type Ⅳ.The production of beta lactamase is mainly mediated by TEM,and just a few strains mediated by ROB-1.To pay attention to the epidemic situation of Hi is of great significance for the selection of antibiotics.