16S rDNA基因序列分析在非哺乳期乳腺炎病原菌检测中的应用价值  

Application of 16S rDNA gene sequencing in the detection of pathogens in non-puerperal mastitis

作　　者：周飞[1] 刘璐[1] 刘丽媛[1] 王斐[1] 于理想[1] 相玉娟 马忠兵[1] 余之刚[1] 

机构地区：[1]山东大学第二医院乳腺外科,山东济南250033

出　　处：《山东大学学报（医学版）》2018年第1期57-61,69,共6页Journal of Shandong University:Health Sciences

基　　金：济南市临床医学科技创新计划(201602146);山东大学第二医院青年基金(Y2014010030)

摘　　要：目的探讨16S r DNA基因序列分析在非哺乳期乳腺炎患者病原菌检测中的应用价值。方法采集33份非哺乳期乳腺炎标本,分别进行传统的细菌镜检、微生物培养与鉴定,并提取其DNA进行PCR扩增及16S rDNA测序分析。结果革兰染色法41.18%(7/17)查见G+细菌,35.29%(6/17)查见G-细菌。微生物培养仅1例阳性(1/5)且呈现两种细菌生长。16S rDNA测序阳性率为100%,其中39.39%(13/33)为混合细菌感染,27.27%(9/33)为假单胞菌属,6.06%(2/33)为kroppenstedtii.棒状杆菌,6.06%(2/33)为屎肠球菌,3.03%(1/33)为坚硬芽孢杆菌,3.03%(1/33)为芽孢八叠球菌,3.03%(1/33)为金黄色葡萄球菌,12.12%(4/33)为宏基因组文库细菌。结论应用16S r DNA测序技术检测非哺乳期乳腺炎病原菌具有简便快速、灵敏度高、特异性强等优点,且可为临床合理选择抗生素提供线索。Objective To investigate the application value of 16S rDNA gene sequencing in detecting pathogens in patients with non-puerperal mastitis. Methods A total of 33 samples of non-puerperal mastiffs were collected. Traditional bacteria microscopy examination, microbial culture and identification were conducted. Meanwhile, the DNA was extracted for PCR amplification and 16S rDNA gene sequencing analysis. Results Bacterial staining group： 17 samples were collected for Gram staining, 41.18% （7/17） of which were Gram positive, and 35.29% （6/17） were Gram negative. Microbial culture group： 5 pus samples were incubated, 1 of which was positive with 2 types of bacteria. 16S rDNA sequencing group： all 33 cases were positive, 39.39% （13/33） of which were mixed bacterial infections, 27.27% （9/33） were Pseudomous, 6.06% （2/33） were kroppenstedtii, corynebacterium, 6.06% （2/33） were faecium. Enterococci, 3.03% （1/33） were Bacillus firmus, 3.03% （1/33） were Sporosarcina, 3.03% （1/33） were Staphylococcus aureus, and 12.12% （4/33） were uncultured bacterium clone. Conclusion The application of 16S rDNA sequencing technique to detect pathogens in non-puerperal mastiffs is safe, simple, highly sensitive and strongly distinctive. It may provide guidance for the rational use of antibiotics.

关 键 词：16S RDNA 非哺乳期乳腺炎 细菌 检测 病因 

分 类 号：R372[医药卫生—病原生物学]