Molecular Characteristics of Neisseria meningitidis Isolated during an Outbreak in a Jail: Association with the Spread and Distribution of ST-4821 Complex Serogroup C Clone in China

Objective To characterize the meningococcal strains isolated from cases and close contacts with meningococcal disease associated with an outbreak in a jail in May 2010 by investigating the national distribution of hyperinvasive ST-4821 serogroup C clone associated with this outbreak. Methods The cases were described based on the clinical symptoms and laboratory results. Pharyngeal swabs were cultured for N. meningitidis from men in the jail. Meningococcal isolates were identified by serogrouping, pulsed-field gel electrophoresis （PFGE） and multilocus sequence typing （MLST）, respectively. Four hundred and sixteen serogroup C N. meningitidis strains were collected from 27 provinces between 2003 and 2010 for a nationwide survey and analyzed by PFGE and MLST. Results Three persons in a jail system were infected with invasive N. meningitidis serogroup C. All isolates tested had matching PFGE patterns and belonged to the multilocus sequence type （ST） 4821 clonal complex. All 47 N. meningitidis strains were identified from the pharyngeal swabs of 166 peoples in the jail, and 26 of them belonged to ST-4821 serogroup C clone, and 90.14% （375/416） serogroup C strains identified in the nationwide survey belonged to the ST-4821 complex. The ST-4821 serogroup C clone was spread nationwide, distributed in 24 provinces, especially in eastern provinces between 2003 and 2010. Conclusion Endemic transmission and carriage rate of ST-4821 serogroup C clone are high in this jail system. The ST-4821 serogroup C clone is spreading in China and nationwide distributed despite the existence of some effective vaccines.

Proteome Analysis of Neisseria meningitidis Serogroup Strains C Associated with Outbreaks in China

Objective During 2003-2005, an outbreak of meningitis due to Neisseria meningitidis serogroup C occurred in China. With the aim to find strain clues result in the final epidemics, the ancestral strain 053442, a clinical isolate, and a carrier strain 053426 with different gene type were analyzed. Methods Clinical strain 053442 and carrier strain 053426 were cultured on GC agar plates under the same condition. Two-dimensional electrophoresis was performed using the pH 3–10 nonlinear IPG strips of 24 cm length, and all the protein spots were identified by matrix-assisted laser desorption/ionization time of flight spectrometry. Results 502 and 380 protein spots were identified in 053426 and 053442 respectively, relating to 266 and 202 different genes covering a wide range of cellular functions. The express volume and number of proteins involved in energy metabolism, protein synthesis and amino acid biosynthesis in 053426 were higher than in 053442. Virulence factor Opa, Opc and a series of proteins involved in pilus assembly and retraction were identified in 053442, which appear to be of primary importance in colonization and invasion of human cells. Compared to 053442, virulence protein species were less in 053426, with lower express volumes too. No Opa and Opc were detected in 053426. Conclusion The different protein expression profiles of the clinical strain 053442 and carrier strain 053426 in the present study provide some clues of the different pathogenicity of the two strains, which may account for result in the final epidemics.

Excellent outcome of primary Neisseria meningitidis keratoconjunctivitis

Infectious conjunctivitis is a very common presentation to medical professional and ophthalmologist all over the world.Although its typically self-limiting and treatable in almost all of the cases,but we need to be aware of the rare and potentially life threatening if the cause is not promptly identified and treated accordingly.In our case report,we highlighted the rare case of Neisseria meningitidis as a primary cause of keratoconjunctivitis.Neisseria meningitidis is a rare etiology of keratoconjunctivitis and its ocular presentations are quite similar with other bacterial or viral infection.The infection may potentially fatal if systemic invasion occurred,however with immediate and proper treatment the outcome is satisfactory.Early diagnosis and proper antibiotic treatment are critical to prevent systemic spread of the infection.Public health intervention is needed to prevent outbreak of the disease.

Antimicrobial susceptibility and serotypes of Neisseria meningitidis and Streptococcus pneumoniae in Sri Lanka:Experience from the National Reference Laboratory

Objective:To determine the antimicrobial susceptibility and serotypes of Neisseria(N.)meningitidis and Streptococcus(S.)pneumoniae in Sri Lankan patients.Methods:We retrospectively analyzed 11 blood culture specimens from suspected patients with invasive meningococcal disease and 26 S.pneumoniae clinical isolates.We tested 6 antimicrobials against N.meningitidis and 12 antimicrobials against S.pneumoniae.Meningococcal serogroup was determined by realtime PCR and Quellung serotyping was used for pneumococcal analysis.Results:N.meningitidis serogroup B was the most common in this study.Intermediate-susceptibility to penicillin was seen in 75.0%(6/8)of strains.Susceptibility to ciprofloxacin,levofloxacin and cotrimoxazole was 62.5%(5/8),62.5%(5/8)and 87.5%(7/8),respectively.Excellent susceptibility was seen in cefotaxime and meropenem.In S.pneumoniae,the most common serotype was 19F in both invasive and non-invasive pneumococcal diseases.The majority of strains showed multidrug resistance.Penicillin non-susceptibility in non-meningeal strains were 13.6%and all meningeal strains were penicillin resistant.Erythromycin was highly resistant in both groups.Amoxicillin showed excellent susceptibility in non-invasive pneumococcal diseases strains.Linezolid,levofloxacin and vancomycin showed 100.0%susceptibility in all pneumococcal isolates.Conclusions:Implementation of vaccines should be considered,especially for children and high-risk populations.This may contribute to reducing pneumococcal and meningococcal invasive disease burden and help prevent emergence of antimicrobial resistant strains.

Surface adhesion and host response as pathogenicity factors of Neisseria meningitidis

Neisseria meningitidis(N.meningitidis) is an exclusively human pathogen that has been identified in 10%-35% of the adult population and in 5.9% of the child population.Despite the high prevalence of carriers of N.meningitidis,it only occasionally causes meningococcal disease in the context of endemic disease,in certain geographic areas or in isolated epidemic outbreaks.After the N.meningitidis genome is described,progress has been made toward understanding the pathogenic mechanisms of the bacteria,although some aspects concerning its interaction with the environment and the host remain unclear.Some studies have reported that oxidative stress in the environment can modify the surface characteristics of N.meningitidis,increasing its adhesive properties and favouring an asymptomaticcarrier state.The antigenic structure of N.meningitidis can be modified by its importing genetic material from other bacteria in its ecological niche.Some structures of lipopolysaccharides help it to evade the immune response,and these are observed more frequently in N.meningitidis isolated from blood than in healthy nasopharyngeal carriers.There is evidence that pili and capsule are downregulated upon contact with target cells.This paper reviews current knowledge on hostenvironment-bacteria mechanisms and interactions,with the aim of contributing to our understanding of the pathogenic mechanisms of N.meningitidis.

Meningitis Outbreak Caused by Neisseria meningitidis Serogroup C ST 10217 in 2019 in Diapaga, Burkina Faso

Introduction: Meningitis caused by Neisseria meningitidis constitutes a burden for the countries in the meningitis belt of sub-Saharan in general and particularly for Burkina Faso. In 2019 the Diapaga health district experienced a meningitis epidemic due to N. meningitidis serogroup C. Methods: This is a cross-sectional study with a descriptive aim in the health district of Diapaga where all cases of meningitis were included in this work. Rapid diagnostic tests (RDTs), culture as well as real-time PCR were used for the biological analysis of cerebral spinal fluid (CSF) samples. Results: Of 155 CSF samples analysed, 42% (65/155) were tested positve. Of them, N. meningitidis C accounted for 83% of all positive cases. Likewise, all thirteen (13) NmC strains were susceptible to oxacillin, ceftriaxone, penicillin and chloramphenicol. All strains of NmC belonged to the sequence type (ST) 10 217 and to the clonal complex (CC) 10 217. These CCs belonged to the same variant PorA type: P1.21-15.16;FetA type: F1-7;PorB type: 3-463. Conclusion: Burkina Faso had known an epidemic of meningitis caused by NmC in 2019 in the health district of Diapaga. This outbreak was contained in time due to the performance of the epidemiological surveillance system which made it possible to investigate on time and introduce the vaccine against the pathogen NmC.

南通市首例B群流行性脑脊髓膜炎病例报道

目的:对南通市首例B群流行性脑脊髓膜炎(以下简称流脑)病例进行报告分析。方法:对2022年2月南通大学附属医院报告的1例实验室确诊流脑病例开展流行病学调查,采集病例血液和脑脊液标本进行脑膜炎奈瑟菌分离培养、血清群PCR鉴定及药物敏感性试验。结果:该病例为26岁女性,自述既往接种过流脑疫苗,接种记录不详,无流脑病例接触史,无续发病例出现。从该病例的血液标本中分离培养出脑膜炎奈瑟菌,经鉴定为脑膜炎奈瑟菌B群;菌株对氨苄西林、头孢噻肟、氯霉素、米诺环素、利福平、左氧氟沙星敏感,对复方新诺明耐药。结论:该病例为南通市报告的首例B群流脑,提示南通市流行菌群可能发生变迁,B群流脑有流行风险,应加强流脑病原学监测。

A first meningococcal meningitis case caused by serogroup X Neisseria meningitidis strains in China

Neisseria meningitidis is the leading cause of bacterial meningitis and classified into 13serogroups based on the immunological reactivity of the capsular polysaccharide.1 Serogroups A, B, C, W135 and Y are the most common causes of meningitis.2 Among them, serogroup A and C are the major causes of epidemics in Africa and Asia.2 Most of the epidemic outbreaks of meningococcal meningitis are caused by serogroup A Neisseria meningitidis strain from the 1950s to the 1980s in China.3 During the years 2003 and 2005, a new sequence type （ST-4821） of serogroup C was identified in the Anhui and 11 other provinces of China.4

Molecular characterizations of serogroup B Neisseria meningitidis strains circulating in Beijing

Neisseria meningitidis （N. meningitidis） is classified into 13 serogroups based on the immunological reactivity of the capsular polysaccharide.Serogourp-s A,B and C are responsible for over 90% of meningococcal disease.2 In developed countries, endemic disease is generally caused by serogroups B and C.

脑膜炎奈瑟菌致老年糖尿病患者感染性眼内炎1例

1例74岁的老年男性糖尿病患者,因右眼红肿胀痛入院,临床上通过患者右眼前房和玻璃体腔抽出液行涂片培养和宏基因组二代测序(mNGS),最终通过mNGS诊断为脑膜炎奈瑟菌引起的感染性眼内炎,经手术治疗和及时有效的抗生素治疗后,患者右眼感染明显控制,眼内炎好转出院。

A Novel Interpretation of Structural Dot Plots of Genomes Derived from the Analysis of Two Strains of Neisseria meningitidis

Neisseria meningitidis is the agent of invasive meningococcal disease, including cerebral meningitis and septicemia. Because the diseases caused by different clonal groups （sequence types） have their own epidemiological characteristics, it is important to understand the differences among the genomes of the N. meningitidis clonal groups. To this end, a novel interpretation of a structural dot plot of genomes was devised and applied; exact nu- cleotide matches between the genomes ofN. meningitidis serogroup A strain Z2491 and serogroup B strain MC58 were identified, leading to the specification of various structural regions. Known and putative virulence genes for each N. meningitidis strain were then classified into these regions. We found that virulence genes of MC58 tend more to the translocated regions （chromosomal segments in new sequence contexts） than do those of Z2491, notably tending towards the interface between one of the translocated regions and the collinear region. Within the col- linear region, virulence genes tend to occur within 16 kb of gaps in the exact matches. Verification of these tendencies using genes clustered in the cps locus was sufficiently supportive to suggest that these tendencies can be used to focus the search for and understanding of virulence genes and mechanisms of pathogenicity in these two organisms.

49株脑膜炎奈瑟菌体外抗菌药物敏感性检测

目的了解2005-2006年中国脑膜奈瑟菌分离株(Neisseria meningitides,Nm)的抗菌药物敏感性变化趋势,为选择抗菌药物对流行性脑脊髓膜炎(流脑)患者进行治疗提供参考。方法采用肉汤稀释检测最低抑菌浓度(MICs)方法、药敏纸片扩散(K-B)法和E-test试纸条检测方法对本实验室收集的49株(16株A群、33株C群)2005-2006年分离的患者Nm菌株进行体外抗菌药物敏感性检测。结果16株A群Nm菌株对复方新诺明、四环素、左氧氟沙星、萘啶酸4种抗菌药物耐药,对环丙沙星耐药或中度敏感。33株C群Nm菌株对复方新诺明耐药,31株(93.9%)C群菌株对萘啶酸耐药,20株(60.6%)C群菌株对左氧氟沙星耐药,17株(51.5%)C群菌株对环丙沙星耐药。分别发现对青霉素不敏感的4株A群和1株C群Nm菌株。结论2005-2006年,中国的A群和C群Nm菌株对磺胺类药物和喹诺酮类药物普遍耐药,此两类药物不适合中国流脑的临床用药及人群预防性给药。K-B药敏纸片和肉汤稀释MICs及E-test方法具有较高的一致性。

2005年广东省健康人群流脑免疫水平和带菌状况调查

目的：了解我省健康人群流脑带菌状况和免疫水平，预测流脑发病的趋势，为流脑的防控提供依据。方法：按照中国疾病预防控制中心的要求并结合广东省的实际情况开展流脑监测工作。选择广州市番禺区、茂名化州市、韶关乐昌市、东莞市及汕头市潮阳区作为流脑监测点。各监测点按广东省流脑监测方案要求采集市内0-、5-、10-、15-、25～、35-及45岁以上共7个年龄组人群的咽拭子及静脉血，分别进行健康人群带菌调查及人群抗体水平检测。结果：广东省健康人群A群流脑IgG抗体的总阳性率为78．9％（675／856），C群流脑IgG抗体的总阳性率为23．7％（203／856）；健康人群脑膜炎奈瑟氏菌（简称Nm）的带菌率为0．65％（7／1077）。健康人带菌以B群Nm为主。结论：广东省不同年龄组健康人群A群流脑IgG抗体的阳性率维持在较高水平，而Nm的带菌率也很低，提示大部分人对A群流脑有免疫作用。但我省健康人群C群IgG抗体水平较低，一旦有传染源传人，易引起局部暴发流行。

2005年贵州省健康人群和病例密切接触者流脑病原菌带菌状况调查

目的了解贵州省健康人群流脑病原菌带菌状况,为流脑的防治工作提供科学依据。方法采集健康人群和病例密切接触者的咽拭标本进行培养、鉴定和分析。结果从904份健康人群标本和56份病例密切接触者的咽拭标本中分别培养出17株和6株脑膜炎奈瑟菌,其中19株(82．61%)为 A 群,平均检出率分别为1．88%和10．71%。10～14岁年龄组学生的带菌率最高,为5．56%。从44份病人脑脊液标本培养出5株脑膜炎奈瑟菌,均为 A 群。结论目前贵州省流脑的流行菌群仍然以 A 群为主,高危人群主要以14岁以下学生为主。提示今后贵州省预防控制流脑的重点应该是加强流脑疫苗的预防接种和流脑病原学监测。

江西省C群脑膜炎奈瑟菌药物敏感性及PFGE分子特征分析

目的分析江西省2006-2010年分离的C群脑膜炎奈瑟菌的药物敏感性及分子特征。方法采用E-test试纸条检测方法对38株C群脑膜炎奈瑟菌菌株进行体外抗菌药物敏感性检测,利用脉冲场凝胶电泳技术(Pulsed-Field Gel Electrophoresis,PFGE)进行分子特征分析。结果所有菌株对氯霉素、美洛培南、阿奇霉素、米洛环素、头孢噻肟、头孢曲松和利福平均敏感;15(39.5%)株菌株对青霉素不敏感,18(47.4%)株对氨苄西林不敏感,25(65.8%)株对环丙沙星耐药,23(60.5%)株对左氧氟沙星耐药,11(28.9%)株对复方新诺明不敏感。PFGE分型显示以AH1和AH2型为主,并且同一地区的带型以同一种带型为主。结论江西省2006-2010年分离C群脑膜炎奈瑟菌株对喹诺酮类药物具有较高的耐药性,还有大部分菌对磺胺类和青霉素类药物也不敏感。PFGE分型特征虽然呈多态性,但是以AH1和AH2型为优势克隆型。

槲皮素对奈瑟球菌感染脑膜炎大鼠继发内毒素血症的抑制作用及机制研究

目的:分析槲皮素对奈瑟球菌感染脑膜炎大鼠继发内毒素血症的抑制作用影响。方法:选取SPF级Wistar雄性大鼠60只,随机数字表法将大鼠分成6组,分别是模型组、正常组、阳性对照组、低剂量槲皮素组、中剂量槲皮素组及高剂量槲皮素组,每组各10只,除正常组大鼠外,其他大鼠制备奈瑟球菌感染脑膜炎模型,造模后阳性对照组灌胃2mL剂量为100mg/L的地塞米松药液,低、中、高剂量槲皮素组灌胃2mL剂量为50、100、200mg/L的槲皮素药液,正常组与模型组大鼠灌胃等剂量生理盐水,各组大鼠给药间隔12h,共给药5次。观察各组大鼠给药12、24、36、48及60h时其存活率状况,同时间隔12h检测大鼠体温一次,ELISA法检测血清IL-6、TNF-α、脂多糖、IL-1β、IL-8含量,Western blot检测p65与IκBα蛋白表达,RT-PCR检测脑组织Toll样受体4(TLR4)mRNA、CD68mRNA相对表达量。结果:造模24h内模型组大鼠全部死亡,模型组大鼠存活率低于正常组,中剂量及高剂量槲皮素组大鼠存活率优于模型组、阳性对照组及低剂量槲皮素组,差异有统计学意义(P<0.05);模型组大鼠血清IL-6、TNF-α、IL-1β及IL-8含量显著高于正常组(P<0.05),其他各给药组血清IL-6、TNF-α、IL-1β及IL-8含量显著低于模型组。其中,中、高剂量槲皮素组大鼠降低显著,和模型组、阳性对照组对比差异有统计学意义(P<0.05)。模型组大鼠IκBα蛋白表达比正常组下降,p65蛋白表达比正常组上升,中、高剂量槲皮素组大鼠IκBα蛋白表达比模型组、阳性对照组上升,p65蛋白比模型组、阳性对照组下降,差异均有统计学意义(P<0.05)。结论:槲皮素可抑制奈瑟球菌感染脑膜炎大鼠机体NF-κB炎症通路,降低血清炎性因子含量,提高大鼠存活率。

江西省2005-2015年脑膜炎奈瑟菌血清分型和分子分型分析

目的对江西省2005-2015年分离的脑膜炎奈瑟菌(Neisseriameningitidis,Nm)进行血清分型和分子分型研究,揭示菌群特征和变迁情况。方法对2005-2015年分离的200株脑膜炎奈瑟菌采用血清凝集的方法鉴定血清群,对引起流脑流行的血清群进行脉冲场凝胶电泳(Pulsed Field Gel Electrophoresis,PFGE)和多位点序列分型(Multilocus Sequence Typing,MLST),使用BioNumerics软件构建最小生成树。分析不同来源、不同年份分离株的血清型和分子型别以及变迁情况。结果 200株脑膜炎奈瑟菌中,有168株可鉴定为已知血清群,包括C群(101株)、B群(50株)、A群(12株)、W135群(2株)、Y群(2株)和X群(1株),另外32株为不可分群菌株。临床病人、密切接触者和健康人群分离株中分别有100%、86.0%和28.9%的菌株为C群。针对健康人群分离株,2005年、2007年、2008年、2012年分离的菌株中,C群分别占1.9%、58.8%、81.5%和0%。95株C群菌株分为14个PFGE型,其中NMNh.JX0001和NMNh.JX0002为优势带型,分别包含46和30株菌。NMNh.JX0001型菌株在2005-2006年、2007-2008年、2013-2014年3个时间段在C群菌株中所占比例分别为97.2%、28.6%、10.0%;而NMNh.JX0002型菌株在2005-2006年、2007-2008年、2013-2014年3个时间段在C群菌株中所占比例分别为2.8%、57.1%、90.0%。挑选优势带型以及相似带型的33株菌株进行MLST分型,均为ST-4821型。结论江西省2005-2015年的流脑病例均为C群菌株引起。在健康人群中存在C群脑膜炎奈瑟菌。虽然近10年流行的C群菌株MLST序列型未发生改变,但是PFGE型别发生了变迁,提示菌株基因组存在微进化。PFGE型别的变迁有可能导致流行模式发生改变,所以有必要开展持续监测,密切关注新型菌株在人群中的传播扩散以及引起的流脑病例。

脉冲场凝胶电泳技术用于脑膜炎奈瑟菌的研究

目的利用脉冲场凝胶电泳技术分析流行性脑脊髓膜炎(简称"流脑")病例及其密切接触者分离出的脑膜炎奈瑟菌(Nm)的同源性以及不同病例分离出Nm的相关性。方法对6例疑似流脑病例的血液和脑脊液标本以及对密切接触者咽拭子进行脑膜炎奈瑟菌的培养分离,分离菌进行氧化酶试验、血清学分群鉴定、糖原利用试验和脉冲场凝胶电泳(PFGE)鉴定试验。结果 6例疑似流脑病例与其密切接触者共分离出21株菌,均为Nm C群,5株为AH 1型,14株为AH 2型,2株为AH 47型。结论流脑病例分离出的Nm与其密切接触者分离出的Nm完全同源,其中4例病例病原菌来自同一菌株。

深圳市2005～2006年健康人群脑膜炎奈瑟菌带菌调查

目的了解2005—2006年深圳市健康人群脑膜炎奈瑟菌（Nm）的带菌状况，为制定有效的防制方案提供依据。方法将健康人群分为0-、5-、10-、15-、25-、35-和45-岁以上7个年龄组，采集咽拭子，进行Nm培养分离和鉴定，用血清学诊断和PCR方法分群。结果2005—2006年深圳市健康人群Nm带菌率为4．4％（55／1255），其中B群49株（89．1％），c群4株（7．3％），29E2株（3．6％）。在两次流脑疫情中各分离到1株B群Nm。结论2005—2006年深圳市健康人群主要携带B群Nm，未检出A群Nm，A群流脑传染源较少。

A群奈瑟氏脑膜炎球菌荚膜多糖-破伤风类毒素结合疫苗的制备及其免疫原性研究

采用溴化氰(CNBr)活化多糖,以无水己二酸二肼(ADH)作为连接剂,1乙基13(3二甲基氨基丙基)碳化二亚胺(EDAC)为偶联剂制备A群奈瑟氏脑膜炎球菌荚膜多糖(GAMP)与破伤风类毒素(TT)的结合物,经皮下免疫NIH小鼠,用ELISA检测小鼠血清中抗GAMP及抗载体蛋白的IgG抗体水平。用补体介导的体外杀菌试验检测血清中GAMP抗体的杀菌活性。结果显示,实验中制备的多糖衍生物和多糖蛋白质结合物都具有GAMP抗原特异活性。结合物免疫小鼠后可诱生比多糖单独免疫更高水平的GAMP血清IgG抗体,并能形成免疫记忆,产生再次应答。结合物免疫小鼠所诱生的血清GAMP抗体较之多糖组具有更强的体外杀菌活性。表明此方法制备的结合物可获得优于多糖的、稳定的特异免疫原性。