PorA Typing Research Articles

Phenotypic, genotypic and proteomic variations between poor and robust colonizing Campylobacter jejuni strains

Campylobacter jejuni is one of the major causes of bacterial gastrointestinal disease in humans worldwide. This foodborne pathogen colonizes the intestinal tracts of chickens, and consumption of chicken and poultry products is identified as a common route of transmission. We analyzed two C. jejuni strains after oral challenge with 105 CFU/ml of C. jejuni per chick; one strain was a robust colonizer (A74/C) and the other a poor colonizer (A74/O). We also found extensive phenotypic differences in growth rate, biofilm production, and in vitro adherence, invasion, intracellular survival, and transcytosis. Strains A74/C and A74/O were genotypically similar with respect to their whole genome alignment, core genome, and ribosomal MLST, MLST, flaA, porA, and PFGE typing. The global proteomes of the two congenic strains were quantitatively analyzed by ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) and 618 and 453 proteins were identified from A74/C and A74/O isolates, respectively. Cluster of Orthologous Groups (COG) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses showed that carbon metabolism and motility proteins were distinctively overexpressed in strain A74/C. The robust colonizer also exhibited a unique proteome profile characterized by significantly increased expression of proteins linked to adhesion, invasion, chemotaxis, energy, protein synthesis, heat shock proteins, iron regulation, two-component regulatory systems, and multidrug efflux pump. Our study underlines phenotypic, genotypic, and proteomic variations of the poor and robust colonizing C. jejuni strains, suggesting that several factors may contribute to mediating the different colonization potentials of the isogenic isolates.

Australian Meningococcal Surveillance Programme Annual Report, 2022.

In Australia, both probable and laboratory-confirmed cases of invasive meningococcal disease (IMD) are reported to the National Notifiable Diseases Surveillance System (NNDSS). Compared to 2021, the number of IMD notifications in 2022 increased by 81% to 127, alongside the easing of COVID-19 containment measures. Laboratory confirmation occurred in 95% of these cases, with 51% (62/121) diagnosed by bacterial culture and 49% (59/121) by nucleic acid amplification testing. The serogroup was determined for 97% of laboratory-confirmed cases (117/121): serogroup B (MenB) accounted for 83% of infections (100/121); MenW for 4% (5/121); MenY for 10% (12/121); no infections were attributed to MenC disease. Fine typing was available on 67% of the cases for which the serogroup was determined (78/117). In MenB isolates, 27 porA types were detected, the most prevalent of which were P1.7-2,4 (18%;11/62), P1.22,14 (15%; 9/62), P1.18-1,34 (10%; 6/62) and P1.7,16-26 (10%; 6/62). All five MenW infections identified as porA type P1.5,2 with different MLST sequence types (ST): 11, 574, 1287, 12351, 13135 all belonging to clonal complex 11, the hypervirulent strain reported in outbreaks in Australia and overseas. In MenY, the predominant porA type was P1.5-1,10-1 (73%; 8/11), ST 1655 and from clonal complex 23. Children less than 5 years of age and people aged 15-19 years were overrepresented with IMD notifications, accounting for 22% (27/121) and 23% (28/121) of laboratory-confirmed cases respectively. Fifteen percent of laboratory-confirmed notifications (18/121) were in persons aged 45-64 years. MenB infections were detected in all age groups but predominated in persons aged 15-19 years (93% of IMD in this age group; 26/28) and comprised 89% (24/27) of infections in children aged less than 5 years. MenW infections were markedly reduced in 2022, accounting for two IMD detections in children 1-4 years (2/16) and sporadic detections in other older age groups. MenY infections were largely detected in adults aged 45-64 years, accounting for 28% of IMD in this age group (5/18). All 62 cultured IMD isolates had antimicrobial susceptibility testing performed. Minimum inhibitory concentration (MIC) values were categorised using Clinical Laboratory Standards Institute (CLSI) interpretative criteria: 5% (3/62) were defined as penicillin resistant (MIC value ≥ 0.5 mg/L); 71% (44/62) had intermediate susceptibility to penicillin (MIC values 0.125 and 0.25 mg/L) and 24% (15/62) were susceptible to penicillin. All isolates were susceptible to ceftriaxone, ciprofloxacin and rifampicin.

Meningitis Outbreak Caused by <i>Neisseria meningitidis</i> 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.

Australian Meningococcal Surveillance Programme Annual Report, 2021.

Invasive meningococcal disease (IMD) is a notifiable disease in Australia, with both probable and laboratory-confirmed cases of IMD reportable to the National Notifiable Diseases Surveillance System (NNDSS). In 2021, there were 74 notifications of IMD made, the lowest number recorded since 1991 when records began. Ninety-one percent of notified cases (67/74) were laboratory confirmed, with 69% of laboratory-confirmed cases (46/67) diagnosed by bacterial culture and 31% (21/67) by nucleic acid amplification testing. The serogroup was determined for 63/67 laboratory-confirmed cases (94%): serogroup B (MenB) accounted for 52% of infections (35/67); MenW for 22% (15/67); MenY for 19% (13/67); there were no infections attributed to MenC. Fine typing was available on 37/67 (55%) where the serogroup was determined. The greatest variability was in MenB, with nine different porA types represented. All MenW infections belonged to a single porA type (P1.5,2) with five different MLST sequence types represented: 11, 574, 1287, 12351, 13135; all belonged to clonal complex 11, the hypervirulent strain reported in recent outbreaks in Australia and overseas. All MenY were from the same porA antigen type, P1.5-1,10-1: MLST sequence type 1655; clonal complex 23. Peaks occurred in children less than 5 years, reaching 24% (16/67) of reported cases, and in those aged 15-19 years reaching 16% (11/67) of reported cases. It is notable that 15% (10/67) of notifications were in persons aged 45-64 years, and an equivalent proportion (15%; 10/67) in adults aged 65 years and above. MenB infections predominated in persons aged 15-19 years (100%, 11/11), and comprised 56% (9/16) of infections in children aged less than 5 years. By contrast, Men W infections accounted for half (5/10) of IMD detections in infants less than 1 year, and 30% (3/10) of infections in persons aged 45-65 years. MenY infections predominated in adults aged 65 years and greater, resulting in 70% (7/10) of IMD in this age group. All 46 IMD isolates had antimicrobial susceptibility testing performed. Minimum inhibitory concentration (MIC) values were categorised using Clinical Laboratory Standards Institute (CLSI) interpretative criteria: 13% (6/46) were defined as penicillin resistant (MIC value, ≥ 0.5 mg/L); 59% (27/46) had intermediate susceptibility to penicillin (MIC values, 0.125 and 0.25 mg/L) and 28% (13/46) were susceptible to penicillin. All isolates were susceptible to ceftriaxone and rifampicin. A single MenB IMD isolate from New South Wales exhibited ciprofloxacin resistance (MIC value, 0.125 mg/L).

Zoonotic Transmission of Campylobacter jejuni to Caretakers From Sick Pen Calves Carrying a Mixed Population of Strains With and Without Guillain Barré Syndrome-Associated Lipooligosaccharide Loci.

Campylobacter jejuni causes foodborne gastroenteritis and may trigger acute autoimmune sequelae including Guillain Barré Syndrome. Onset of neuromuscular paralysis is associated with exposure to C. jejuni lipooligosaccharide (LOS) classes A, B, C, D, and E that mimic and evoke antibodies against gangliosides on myelin and axons of peripheral nerves. Family members managing a Michigan dairy operation reported recurring C. jejuni gastroenteritis. Because dairy cattle are known to shed C. jejuni, we hypothesized that calves in the sick pen were the source of human infections. Fecal samples obtained from twenty-five calves, one dog, and one asymptomatic family member were cultured for Campylobacter. C. jejuni isolates were obtained from thirteen calves and the family member: C. coli from two calves, and C. hyointestinalis from two calves. Some calves had diarrhea; most were clinically normal. Typing of lipooligosaccharide biosynthetic loci showed that eight calf C. jejuni isolates fell into classes A, B, and C. Two calf isolates and the human isolate possessed LOS class E, associated mainly with enteric disease and rarely with Guillain Barré Syndrome. Multi-locus sequence typing, porA and flaA typing, and whole genome comparisons of the thirteen C. jejuni isolates indicated that the three LOS class E strains that included the human isolate were closely related, indicating zoonotic transmission. Whole-genome comparisons revealed that isolates differed in virulence gene content, particularly in loci encoding biosynthesis of surface structures. Family members experienced diarrheal illness repeatedly over 2 years, yet none experienced GBS despite exposure to calves carrying invasive C. jejuni with LOS known to elicit antiganglioside autoantibodies.

Estimated strain coverage of serogroup B meningococcal vaccines: A retrospective study for disease and carrier strains in Greece (2010–2017)

Invasive meningococcal disease (IMD) is associated with high case fatality rates and long-term sequelae among survivors. Meningococci belonging to six serogroups (A, B, C, W, X, and Y) cause nearly all IMD worldwide, with serogroup B meningococci (MenB) the predominant cause in many European countries, including Greece (~80% of all IMD). In the absence of protein-conjugate polysaccharide MenB vaccines, two protein-based vaccines are available to prevent MenB IMD in Greece: 4CMenB (Bexsero™, GlaxoSmithKline), available since 2014; and MenB-FHbp, (Trumenba™, Pfizer), since 2018. This study investigated the potential coverage of MenB vaccines in Greece using 107 MenB specimens, collected from 2010 to 2017 (66 IMD isolates and 41 clinical samples identified solely by non-culture PCR), alongside 6 MenB isolates from a carriage study conducted during 2017–2018. All isolates were characterized by multilocus sequence typing (MLST), PorA, and FetA antigen typing. Whole Genome Sequencing (WGS) was performed on 66 isolates to define the sequences of vaccine components factor H-binding protein (fHbp), Neisserial Heparin Binding Antigen (NHBA), and Neisseria adhesin A (NadA). The expression of fHbp was investigated with flow cytometric meningococcal antigen surface expression (MEASURE) assay. The fHbp gene was present in-frame in all isolates tested by WGS and in 41 MenB clinical samples. All three variant families of fHbp peptides were present, with subfamily B peptides (variant 1) occurring in 69.2% and subfamily A in 30.8% of the samples respectively. Sixty three of 66 (95.5%) MenB isolates expressed sufficient fHbp to be susceptible to bactericidal killing by MenB-fHbp induced antibodies, highlighting its potential to protect against most IMD in Greece.

Cross-reactivity of 4CMenB vaccine-induced antibodies against meningococci belonging to non-B serogroups in Italy

ABSTRACT The four-component meningococcal serogroup B vaccine (4CMenB) contains antigens present in the majority of meningococci causing invasive meningococcal disease (IMD) and may potentially offer protection against strains belonging to non-B serogroups. This study aimed to evaluate the ability of 4CMenB-induced antibodies to kill, in a human serum bactericidal assay (hSBA), non-B meningococci belonging to the main genotypes responsible for IMD in Italy. Meningococci, collected between 2015 and 2017, was characterized for PorA, FetA and sequence type, and for clonal complex. Twenty non-B isolates, representative of the most frequent genotypes, were molecularly characterized for 4CMenB antigens and tested in hSBA with sera from 4CMenB-vaccinated infants and adolescents. Among twenty isolates, eleven were serogroup C, five were Y, two W and two X. All isolates contained genes encoding for fHbp and NHBA antigens and four harbored the NadA full-length encoding gene. Positive hSBA titers were obtained against all serogroup W, X and Y isolates and against five serogroup C isolates. These data show that the 4CMenB vaccine can induce bactericidal antibodies against genetically representative meningococcal W, Y and X strains from Italy. For serogroup C, different susceptibilities to killing were observed for strains with similar antigenic repertoires.

Longitudinal study of meningococcal carriage rates in university entrants living in a dormitory in South Korea.

University students, especially those living in dormitories, are known to have a high risk of invasive meningococcal disease. We performed a longitudinal study to investigate the change in Neisseria meningitidis carriage rates and identify the risk factors for carriage acquisition in university students in South Korea. We recruited university entrants who were admitted to a student dormitory. Pharyngeal swabs were taken from participants at baseline, 1 month, and 3 months, and the subjects completed a questionnaire. Culture and real-time polymerase chain reaction (PCR) for species-specific ctrA and sodC genes were performed. The cultured isolates or PCR-positive samples were further evaluated for epidemiologic characterization using serogrouping, PorA typing, FetA typing, and multilocus sequence typing (MLST). At the first visit, we enrolled 332 participants who were predominantly male (64.2%) with a median age of 19 years. Meningococcal carriage rates increased from 2.7% (95% confidence interval [CI] 0.9–4.4%) at baseline to 6.3% (95% CI 3.4–9.0%) at 1 month and 11.8% (95% CI 7.8–15.6%) at 3 months. Nongroupable isolates accounted for 50.0% of all isolates, with serogroup B being the next most prevalent (24.1%). In the study population, male sex (OR 2.613, 95% CI 1.145–5.961, p = 0.022) and frequent pub or club visits (OR 3.701, 95% CI 1.536–8.919, p = 0.004) were significantly associated with meningococcal carriage. Based on serotype and MLST analyses, six carriers transmitted meningococci to other study participants. N. meningitidis carriage rates among new university entrants who lived in a dormitory significantly increased within the first 3 months of dormitory stay, probably owing to the transmission of identical genotype among students. Based on the risk of meningococcal disease, meningococcal vaccination should be considered for students before dormitory admission.

Epidemiology of invasive meningococcal disease in Greece, 2006-2016.

The present study describes the epidemiology of invasive meningococcal disease (IMD) in Greece for the period 2006-2016. Combined data from notified and laboratory-confirmed IMD cases were obtained from the two involved National Centres (Epidemiology and Reference Laboratory). Laboratory identification and typing was carried out by both conventional (culture) and molecular methods (PCR, MLST, PorA, and FetA typing). A total of 796 IMD cases were notified; of those, 720 (91%) were laboratory confirmed. Overall, a decline on the annual incidence of confirmed cases was observed, ranging from 0.91 (2006) to 0.47 (2016) /100,000. A similar trend was observed in most age groups especially in children 0-4years (7.7 to 2.9/100,000), with the exception of an increase in the incidence rate in adults > 20years (0.21 to 0.32/100,000). The overall case fatality rate was 6.5% (52/796), annual range 2-13%. Among 658 strains which were typed by sero/genogroup, 80% were identified as MenB (annual range 65-92%); however, a decline was observed in MenB incidence from 5.3 (2006) to 2.7 (2016), among infants and toddlers, while MenW (1%), MenY (2%), and MenA (1%) remained low. During the 11years, the annual incidence of IMD declined by 50%, especially in the 0-4-year age group, due mainly to MenB. Continuous surveillance of IMD is important for the development of future vaccination and public health policies.

Carriage meningococcal isolates with capsule null locus dominate among high school students in a non-endemic period, Italy, 2012–2013

Meningococcal disease incidence in Italy remains quite low in the overall population except for infants. Within a study on carriage isolates among high school students we aimed to define: i) the prevalence of carriage isolates, ii) the phenotypic and iii) the molecular features of meningococci by Whole Genome Sequencing (WGS).A total of 1697 pharyngeal samples from undergraduate students (age range 14–19 years) were collected from 2012 to 2013 from six larger cities in Italy. One hundred and twenty culture positive meningococci (7%) were analyzed. Carriage isolates were sent to the National Reference Laboratory for invasive meningococcal disease (IMD) for PCR-based serogroup identification, Multilocus Sequence Typing, PorA and FetA typing. Moreover, factor H binding protein (fHbp), Neisseria Heparin Binding Antigen (NHBA) and Neisserial adhesin A (NadA) were typed. Core genome MLST (cgMLST) was performed on a subsample of 75 carriage isolates.Capsule null locus (cnl) predominated (47%), followed by serogroup B (27%). The antimicrobial susceptibility profile revealed an high prevalence of reduced susceptibility to penicillin G (54%) and a full susceptibility to ceftriaxone, ciprofloxacin and rifampicin. Carriage isolates presented a high genetic diversity: the clonal complexes (ccs) cc1136, cc198 and cc41/44, were the predominant. An high heterogeneity was also observed for PorA and FetA types. The fhbp and nhba genes were identified in all the carriage isolates; only 5% of the carriage isolates presented the nadA gene. The core genome MLST analysis revealed that the majority of the cnl isolates clustered in a distinct group.The evidence gathered during this study provides the estimate of carriage isolates in high school students in a non-epidemic period in Italy that was lower than expected. Moreover, the highest proportion of carriage isolates were cnl and, overall, they were molecular heterogeneous.

Genetic Diversity of Campylobacter Jejuni Isolated from the Poultry Food Chain.

IntroductionCampylobacter jejuni is one of the most frequently reported causes of foodborne bacterial enteric disease worldwide. The main source of these microorganisms is contaminated food, especially of poultry origin. There are several molecular methods for differentiation of Campylobacter isolates at the subgenus level, and one of these is porA-typing based on the sequencing of the major outer-membrane protein (MOMP) encoding gene. The aim of the study was to test the molecular relationship of C. jejuni strains isolated at different points along the poultry food chain and assess the population structure of the isolates.Material and MethodsA total of 451 C. jejuni were used in the study, and a DNA fragment of 630 bp of the MOMP encoding gene was amplified and sequenced.ResultsOne hundred and ten sequence types were identified, with 69 (62.7%) unique to the isolates' origin and 30 not present in the database. The most prevalent nucleotide variant 1 was detected in 37 (8.2%) strains. These isolates were identified in all poultry sources tested, especially in faeces (15 isolates) but also in poultry carcasses and meat (11 isolates in each).ConclusionThe porA typing method was highly discriminative for C. jejuni of poultry origin since the Simpson's diversity index (D) achieved a value of 0.876, indicating considerable diversity in the bacterial population tested. The method may be further used for epidemiological investigation purposes.

Molecular characteristics of serogroup B neisseria meningitidis, China

Objective: To investigate the molecular characteristics of serogroup B neisseria meningitidis in China. Methods: Total of 485 (100 strains isolated from cerebrospinal fluid or blood samples of encephalomyelitis cases, and 385 strains isolated from nasopharynx of healthy carriers) Meningococcal serogroup B (MenB) strains, isolated from 29 provinces of China between 1968 and 2016, were analyzed by multilocus sequence typing (MLST) and PorA typing methods. Further, the genetic diversity of three MenB vaccine proteins, FHbp, NadA and NHBA, were analyzed. Results: The 485 study strains belonged to 270 sequence types (STs), 107 of which (representing 211 strains) could be grouped into ten clonal complexes (CC). CC4821 has been the predominant lineage in China since 2005 (28.7%, n=139). The most common PorA types of MenB strains from invasive meningococcal cases were P1.5-2,2-2 (10.0%, n=10), P1.5-1,2-2 (9.0%, n=9) and P1.5-1,10-4 (9.0%, n=9). Four hundred and twenty one strains had intact fhbp gene; variant 1, 2 and 3 accounted for 12.8% (54 strains), 85.0% (358 strains) and 2.2% (9 strains) respevtively. Ten out of 432 strains (2.3%) contained complete nadA gene. All the 172 strains for which the nhba gene was sequenced had intact gene sequence which corresponded to 68 peptide types. Conclusion: CC4821 was the predominant CC of MenB strains in China; the vaccine proteins were diverse about the sequences. The vaccine proteins should be carefully selected when developing MenB vaccines in China.

Molecular epidemiology and antimicrobial resistance mechanisms of Campylobacter coli from diarrhoeal patients and broiler carcasses in Belgium.

The aim of this study was to better understand the molecular epidemiology of Campylobacter coli isolated from multiple sources in Belgium, by studying the genotypic diversity and antimicrobial resistance phenotypes and resistance mechanisms of 59 C.coli isolates. Isolates from broiler carcasses and human cases were genotyped using multilocus sequence typing (MLST), porA typing, flagellin gene A restriction fragment length polymorphism (flaA-RFLP) typing, and by PCR binary typing (P-BIT). Thirty-two MLST sequence types, 24 flaA types, 31 porA alleles, and 29 P-BIT types were identified among the screened isolates. Some types and alleles were shared among strains recovered from both broiler carcasses and diarrhoeal patients. Both porA typing and MLST revealed a similar discriminatory power (0.969), which was the highest discriminatory power when compared to other methods. Minimum inhibitory concentrations against seven different antibiotics (ciprofloxacin, chloramphenicol, nalidixic acid, streptomycin, tetracycline, gentamicin, and erythromycin) were analysed. Strains were most frequently resistant to tetracycline (81.4%), followed by: ciprofloxacin and nalidixic acid (76.3%); streptomycin (33.9%); erythromycin (27.1%); and chloramphenicol (3.4%). All isolates were sensitive to gentamicin. Multidrug resistance was observed in 24 of 59 C.coli isolates (40.7%). Molecular screening of antimicrobial resistance mechanisms revealed the predominance of the gyrA T86I substitution among ciprofloxacin- and nalidixic acid-resistant isolates, the tet(O) gene among tetracycline-resistant isolates and the 23S rRNA A2075G mutation among erythromycin- resistant isolates. Furthermore, some erythromycin-resistant isolates harboured a diverse array of resistance mechanisms, including the presence of ermB, 23S rRNA A2074G mutation, and point mutations the rplD and rplV ribosomal genes, and the cmeABC multidrug efflux pump genes.

PorA typing of Neisseria meningitidis isolates from Iranian children for vaccine design

PorA typing of Neisseria meningitidis isolates from Iranian children for vaccine design

Predicted strain coverage of a meningococcal multicomponent vaccine (4CMenB) in Portugal.

ObjectiveAlthough the incidence of meningococcal disease has been declining over the past decade in Portugal MenB meningococci is still an important cause of meningitis and sepsis. The aim of this study was to estimate the strain coverage of the 4CMenB vaccine in Portugal in order to support health policies for prevention and control of meningococcal disease.MethodsSince 2002 the clinical and laboratory notification of meningococcal disease is mandatory in Portugal. National database includes since then all confirmed cases notified to the reference laboratory or to the Directorate of Health. Strains included in this study were all the invasive MenB isolated from the 1st July 2011 to the 30th June 2015, sent to the reference laboratory. To predict the vaccine strain coverage of the 4CMenB the expression and cross-reactivity of the surface antigens fHbp, NadA, NHBA were assessed by the Meningococcal Antigen Typing System (MATS) whereas PorA typing was performed by sequencing. The presence of at least one antigen with a Relative Potency (RP) greater than its MATS-positive bactericidal threshold RP value or the presence of PorA VR2 = 4 was considered to be predictive for a strain to be covered by the 4CMenB vaccine.ResultsThe estimated 4CMenB strain coverage in Portugal was 67.9%. The percentage of strain coverage in each of the four epidemiological years ranged from 63.9% to 73.7%.Strains covered by one antigen represent 32.1% of the total of isolates, 29.2% of strains were covered by two antigens and 6.6% by three antigens. No strain had all the four antigens. Antigens that most contributed for coverage were NHBA and fHbp.Data from Portugal is in accordance with the MATS predicted strain coverage in five European countries (England and Wales, France, Germany, Italy and Norway) that pointed to 78% coverage for strains collected in the epidemiological year 2007–2008.

Invasive Meningococcal Meningitis Serogroup C Outbreak in Northwest Nigeria, 2015 - Third Consecutive Outbreak of a New Strain.

BACKGROUND: In northwest Nigeria in 2013 and 2014, two sequential, localized outbreaks of meningitis were caused by a new strain of Neisseria meningitidis serogroup C (NmC). In 2015, an outbreak caused by the same novel NmC strain occurred over a wider geographical area, displaying different characteristics to the previous outbreaks. We describe cases treated by Médecins Sans Frontières (MSF) in the 2015 outbreak. METHODS: From February 10 to June 8, 2015, data on cerebrospinal meningitis (CSM) cases and deaths were recorded on standardized line-lists from case management sites supported by MSF. Cerebrospinal fluid (CSF) samples from suspected cases at the beginning of the outbreak and throughout from suspected cases from new geographical areas were tested using rapid Pastorex® latex agglutination to determine causative serogroup. A subset of CSF samples was also inoculated into Trans-Isolate medium for testing by the WHO Collaborating Centre for Reference and Research on Meningococci, Oslo. Reactive vaccination campaigns with meningococcal ACWY polysaccharide vaccine targeted affected administrative wards. RESULTS: A total of 6394 (65 confirmed and 6329 probable) cases of CSM including 321 deaths (case fatality rate: 5.0%) were recorded. The cumulative attack rate was 282 cases per 100,000 population in the wards affected. The outbreak lasted 17 weeks, affecting 1039 villages in 21 local government areas in three states (Kebbi, Sokoto, Niger). Pastorex® tests were NmC positive for 65 (58%) of 113 CSF samples. Of 31 Trans-Isolate medium samples, 26 (84%) tested positive for NmC (14 through culture and 12 through PCR); all had the same rare PorA type P1.21-15,16 as isolates from the 2013 and 2014 outbreaks. All 14 culture-positive samples yielded isolates of the same genotype (ST-10217 PorA type P1.21-15,16 and FetA type F1-7). More than 222,000 targeted individuals were vaccinated relatively early in the outbreak (administrative coverage estimates 98% and 89% in Kebbi and Sokoto, respectively). CONCLUSIONS: The outbreak was the largest caused by NmC documented in Nigeria. Reactive vaccination in both states may have helped curtail the epidemic. A vaccination campaign against NmC with a long-lasting conjugate vaccine should be considered in the region.

Clonal characteristics of invasive Neisseria meningitidis following initiation of an A + C vaccination program in China, 2005–2012

To reveal the population structure of invasive Neisseria meningitidis isolates after the initiation of an A + C vaccination program in China. Multilocus sequence typing (MLST) and PorA typing were used to characterize 238 invasive N. meningitidis isolates collected in China between 2005 and 2012. During this period, sequence type (ST)-5, ST-4821 and ST-11 complexes were dominant among serogroups A, C and W, accounting for 100%, 98.5% and 100% of each serogroup, respectively. P1.20,9, P1.7-2,14 and P1.5,2 were the dominant PorA types of serogroups A, C and W, respectively. Serogroup B showed high genetic diversity with two dominant lineages: ST-4821 complex and ST-5662 subgroup. The population of 238 invasive N. meningitidis isolates was primarily composed of a select group of recognized hypervirulent lineages. Among these clonal complexes, ST-7 serogroup A and ST-11 serogroup W are distributed globally, and other three clones exist only in China.

Molecular and Serological Diversity of Neisseria meningitidis Carrier Strains Isolated from Italian Students Aged 14 to 22 Years

Neisseria meningitidis is an obligate human commensal that commonly colonizes the oropharyngeal mucosa. Carriage is age dependent and very common in young adults. The relationships between carriage and invasive disease are not completely understood. In this work, we performed a longitudinal carrier study in adolescents and young adults (173 subjects). Overall, 32 subjects (18.5%) had results that were positive for meningococcal carriage in at least one visit (average monthly carriage rate, 12.1%). Only five subjects tested positive at all four visits. All meningococcal isolates were characterized by molecular and serological techniques. Multilocus sequence typing, PorA typing, and sequencing of the 4CMenB vaccine antigens were used to assess strain diversity. The majority of positive subjects were colonized by capsule null (34.4%) and capsular group B strains (28.1%), accounting for 23.5% and 29.4% of the total number of isolates, respectively. The fHbp and nhba genes were present in all isolates, while the nadA gene was present in 5% of the isolates. The genetic variability of the 4CMenB vaccine antigens in this collection was relatively high compared with that of other disease-causing strain panels. Indications about the persistence of the carriage state were limited to the time span of the study. All strains isolated from the same subject were identical or cumulated minor changes over time. The expression levels and antigenicities of the 4CMenB vaccine antigens in each strain were analyzed by the meningococcal antigen typing system (MATS), which revealed that expression can change over time in the same individual. Future analysis of antigen variability and expression in carrier strains after the introduction of the MenB vaccine will allow for a definition of its impact on nasopharyngeal/oropharyngeal carriage.

Molecular Characterization of Invasive Neisseria meningitidis Strains Isolated in Chile during 2010–2011

BackgroundWith the upcoming licensure of Outer Membrane Protein-based vaccines against meningococcal disease, data on disease incidence and molecular characteristic of circulating N. meningitidis strains in Latin American countries is needed. Chile is, to date, one of the few countries in the region that has performed this type of work in a comprehensive collection of disease-associated strains from two consecutive years, 2010–2011.MethodsA total of 119 N. meningitidis strains isolated from patients with invasive disease in Chile in 2010–2011 were characterized by the National Reference Laboratory. Serogroup determination, MLST and porA typing were performed.ResultsSerogroup B was predominant in both study years, but W135 experienced a noticeable increase in 2011 compared to 2010. ST-11 complex, ST-41/44 complex ST-32 complex were the most prevalent among the isolates, and were strongly associated with serogroups W135 (ST-11 Complex) and B (ST-41/44 and ST-32 complexes). Likewise, the major porA types detected were strongly associated with these three clonal complexes: P1.5,2 was found exclusively among W135:ST-11 isolates, whereas P1.7, 2–3 was only detected in C:ST-11. ST-41/44 isolates mainly had P1.10-8, and ST-32 complex were associated with a P1.18-8 porA.ConclusionsOur data show disease-associated N. meningitidis circulating in Chile are similar to those found in other parts of the world. The increase on W135:ST-11 isolates observed in 2011 foretold the unusual epidemiological situation experienced in the country in 2012, and MLST data show that this strain is indistinguishable from the one linked to the global Hajj 2000-related outbreak that occurred in 2001. Finally, this work demonstrates the importance of maintaining a strong national surveillance program integrating clinical, epidemiological and laboratory data and incorporating gold standard diagnostic and characterization techniques that allow the data to be compared all over the world.

Fatal Neisseria meningitidis serogroup X sepsis in immunocompromised patients in Spain. Virulence of clinical isolates

Neisseria meningitidis X (NmX) is a minority meningococcus serogroup for which no vaccine is available. Huge meningococcus X outbreaks occurred in West Africa but developed countries have reported only sporadic cases. Two invasive fatal cases are presented. Isolates were characterized by serogrouping (latex agglutination and genogroup X-specific polymerase-chain-reaction), PorA and FetA typing, multilocus sequence typing and pulsed-field gel electrophoresis. Immunocompetent female balm/C mice inoculated intraperitoneally were use to test virulence of invasive and carrier isolates. Until April 2010, NmX was absent among 868 invasive meningococci characterized in the Basque Country in a 20-year period. In April 2010, two fatal NmX: P1.21,16: F5-5/ST750 episodes were detected in unrelated immunodeficient patients. After analysis of 803 meningococcal isolates from asymptomatic carriers obtained between 1988 and 2010, eight NmX isolates were detected. The genotype of the two invasive isolates bore no relation to any of the NmX isolates detected in healthy individuals from the Basque Country or to isolates from outbreaks in Africa. NmX isolates in the north of Spain can cause severe disease in humans, despite their low prevalence. The in vivo animal study showed that virulence of isolates was more closely associated with the genotype than with the serogroup.