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目的对2013年1月北京市1例流行性脑脊髓膜炎(流脑)死亡病例标本进行脑膜炎奈瑟菌检测,了解感染病原及分离菌株的分子流行病学特征。方法对患者脑脊液标本进行细菌分离培养,通过常规细菌学试验对分离菌株进行脑膜炎奈瑟菌鉴定、实时荧光定量-聚合酶链反应(real-time fluorescence quantitative PCR)方法检测患者脑脊液和抗凝血中脑膜炎奈瑟菌核酸,同时对鉴定的菌株进行多位点序列分型(MLST)分析、外膜蛋白PorA分型和FetA分型,以及体外药物敏感试验。结果从患者脑脊液和抗凝血标本中均检测到脑膜炎奈瑟菌种特异基因ctrA和血清群B特异基因siaD。从患者脑脊液中分离到1株可疑阳性菌,经实验确诊为B群脑膜炎奈瑟菌。PorA为P1.20,23-9,FetA为F1-91;多位点序列分型分析显示基因型为ST10051,属于ST4821序列群。该菌株对复方新诺明为中等耐药,对环丙沙星和四环素耐药,对青霉素、头孢噻肟、头孢曲松、左氧氟沙星、美罗培南、氯霉素、利福平等药物均敏感。从密切接触者咽拭子中未分离到脑膜炎奈瑟菌。结论该菌株经鉴定为B:P1.20,23-9:F1-91:ST10051(ST4821序列群)。北京地区人群中已经存在高致病性ST4821克隆群B群脑膜炎奈瑟菌,并引发死亡病例,提示应全面掌握本地区脑膜炎奈瑟菌健康人群携带情况及病例菌株分子流行病学资料,为流脑的预防控制提供实验室数据支撑。
Objective To detect Neisseria meningitides in a case of epidemic cerebrospinal meningitis (meningitis) in Beijing from January 2013 to January 2006, and to understand the molecular epidemiology of infectious pathogens and isolates. Methods The cerebrospinal fluid (CSF) samples of patients were isolated and cultured. Neisseria meningitidis was identified by routine bacteriological tests, and cerebrospinal fluid and anticoagulant blood samples were measured by real-time fluorescence quantitative PCR Neisseria meningitidis nucleic acid. Meanwhile, MLST analysis, PorA typing and FetA typing of the outer membrane protein, and in vitro drug sensitivity test were performed on the identified strains. Results Neisseria meningitidis-specific gene ctrA and serogroup B-specific gene siaD were detected in both cerebrospinal fluid and anticoagulant samples. From the patient’s cerebrospinal fluid was isolated from a suspicious positive bacteria, which was diagnosed as Neisseria meningitides group B. PorA was P1.20,23-9 and FetA was F1-91. Multiple loci sequence analysis revealed that the genotype was ST10051, belonging to the ST4821 sequence group. The strain was moderately resistant to cotrimoxazole, resistant to ciprofloxacin and tetracycline, and sensitive to penicillin, cefotaxime, ceftriaxone, levofloxacin, meropenem, chloramphenicol and rifampicin. Neisseria meningitidis was not isolated from throat swabs in close contacts. Conclusion This strain was identified as B: P1.20, 23-9: F1-91: ST10051 (ST4821 sequence group). Neisseria meningitidis serogroup B of highly pathogenic ST4821 population has already existed in Beijing population and caused deaths, suggesting that the epidemiological data of healthy individuals of Neisseria meningitidis in this area should be fully grasped, Laboratory data support for the prevention and control of meningitis.