ABSTRACT
Introduction. Colonization by Neisseria meningitidis is the pre-requisite for the development of disease. We present the findings of a cross-sectional investigation onto the oropharyngeal carriage of N. meningitidis and Neisseria species in the population aged 3 to 21 in Paraguay.Aim. Carriage prevalence by age groups, risk factors associated with carriage, and phenotypic and genotypic characteristics of strains are described.Methodology. We collected 2011 oropharyngeal swabs from consenting participants aged 3-21 years. Infants were recruited at immunization clinics, and older children and young adults were identified at schools and universities. A single oropharyngeal swab was collected and processed for the identification and isolation of Neisseria. Additionally, participants, or their legal guardian if these were minors, were requested to fill a standardized questionnaire.Results. N. meningitidis was isolated in 42/2011 (2.1 %) participants, while other Neisseria spp. were identified in 306/2011 (15.2 %) subjects: N. cinerea and N. lactamica were identified in 39/2011 (1.9 %) and 43/2011 (2.2 %), respectively. Meningococcal strains belonged to ten different clonal complexes, of which six are associated with invasive disease (ST-32/ET5 complex, ST-11/ET37 complex, ST-103 complex, ST-167 complex, ST-35 complex and ST-41/44 complex/lineage 3).Conclusion. Prevalence of N. meningitidis carriage was low compared to that reported from other settings, however, the overall carriage of Neisseria spp. (including N. meningitidis) was comparable to meningococcal carriage prevalence reported in the literature. This study is the first of its kind conducted in Paraguay, and one of the few known in the Southern Cone of Latin America.
Subject(s)
Carrier State/epidemiology , Neisseria meningitidis/isolation & purification , Neisseria/isolation & purification , Oropharynx/microbiology , Adolescent , Child , Child, Preschool , Cross-Sectional Studies , Female , Humans , Male , Neisseria/classification , Paraguay , Young AdultABSTRACT
OBJECTIVE: To evaluate the impact of a collaborative programme for the early recognition and management of patients admitted with sepsis in the northwest of England. SETTING: 14 hospitals in the northwest of England. INTERVENTION: A quality improvement programme (Advancing Quality (AQ) Sepsis) that promoted a sepsis care bundle including time-based recording of early warning scores, documenting systemic inflammatory response syndrome criteria and suspected source of infection, taking of blood cultures, measuring serum lactate levels, administration of intravenous antibiotics, administration of oxygen, fluid resuscitation, measurement of fluid balance and senior review. MAIN OUTCOME MEASURES: Inpatient mortality, 30-day readmission rates and duration of hospital ≥10 days. RESULTS: Data for 7776 patients were included in this study between 1 July 2014 and 29 December 2015. Participation in the AQ Sepsis programme was associated with a reduction in readmissions within 30 days (OR 0.81 (0.69-0.95)) and hospital stays over 10 days (OR 0.69 (0.60-0.78)). However, there was no reduction in mortality. Administration of a second litre of intravenous fluid within 2 hours, oxygen therapy and review by a senior clinician were associated with increased mortality. Starting a fluid balance chart within 4 hours was the only clinical process measure that did not affect mortality. Taking a blood culture sample, administering antibiotic therapy and measuring serum lactate within 3 hours of hospital arrival were all associated with reduced mortality (OR 0.69 (0.59-0.81), OR 0.77 (0.67-0.89) and OR 0.64 (0.54-0.77), respectively) and shorter hospitalisations (OR 0.58 (0.49-0.69), OR0.81 (0.70-0.94) and OR 0.54 (0.45-0.66), respectively). However, none of these measures had an impact on the risk of readmission to hospital within 30 days. CONCLUSIONS: The AQ Sepsis collaborative in northwest of England improved readmission and length of stay for patients admitted with sepsis but did not affect mortality. Further cost-effectiveness evaluation of the programme is needed.
Subject(s)
Patient Care Bundles , Quality Improvement , Sepsis/diagnosis , Sepsis/therapy , Adolescent , Adult , Aged , Aged, 80 and over , Early Diagnosis , England/epidemiology , Female , Hospital Mortality , Humans , Length of Stay/statistics & numerical data , Male , Middle Aged , Patient Readmission/statistics & numerical data , Retrospective Studies , Sepsis/mortalityABSTRACT
OBJECTIVE: To evaluate the impact of integrating a general practitioner (GP) into a tertiary paediatric emergency department (ED) on admissions, waiting times and antibiotic prescriptions. DESIGN: Retrospective cohort study. SETTING: Alder Hey Children's NHS Foundation Trust, a tertiary paediatric hospital in Liverpool, UK. PARTICIPANTS: From October 2014, a GP was colocated within the ED, from 14:00 to 22:00 hours, 7 days a week. Children triaged green on the Manchester Triage System without any comorbidities were classed as 'GP appropriate'. The natural experiment compared patients triaged as 'GP appropriate' and able to be seen by a GP between 14:00 and 22:00 hours (GP group) to patients triaged as 'GP appropriate' seen outside of the hours when a GP was available (ED group). Intention-to-treat (ITT) analysis was used to assess the main outcomes. RESULTS: 5223 patients were designated as 'GP appropriate'-18.2% of the total attendances to the ED over the study period. There were 2821 (54%) in the GP group and 2402 (46%) in the ED group. The median duration of stay in the ED was 94 min (IQR 63-141) for the GP group compared with 113 min (IQR 70-167) for the ED group (p<0.0005). Using the ITT analysis equivalent, we demonstrated that the GP group were less likely to: be admitted to hospital (2.2% vs 6.5%, OR 0.32, 95% CI 0.24 to 0.44), wait longer than 4 hours (2.3% vs 5.1%, OR 0.45, 95% CI 0.33 to 0.61) or leave before being seen (3.1% vs 5.7%, OR 0.53, 95% CI 0.41 to 0.70), but more likely to receive antibiotics (26.1% vs 20.5%, OR 1.37, 95% CI 1.10 to 1.56). Sensitivity analyses yielded similar results. CONCLUSIONS: Introducing a GP to a paediatric ED service can significantly reduce waiting times and admissions, but may lead to more antibiotic prescribing. This study demonstrates a novel, potentially more efficient ED care pathway in the current context of rising demand for children's emergency services.
Subject(s)
Emergency Service, Hospital/organization & administration , General Practitioners/statistics & numerical data , Patient Admission/statistics & numerical data , Referral and Consultation/statistics & numerical data , Triage/statistics & numerical data , Adolescent , Anti-Bacterial Agents/therapeutic use , Checklist , Child , Child, Preschool , Drug Utilization , England , Female , Hospitals, Pediatric , Humans , Infant , Intention to Treat Analysis , Length of Stay , Male , Retrospective Studies , Tertiary Care Centers , United Kingdom , Waiting ListsABSTRACT
BACKGROUND: The lack of information regarding the burden of acute bacterial meningitis in Latin America leads to a reduction in the estimated incidence rates of the disease, and impairs public health decisions on the use and follow-up of preventive interventions, particularly, the evaluation of existing vaccination policies. The use of the real-time PCR in diagnostic routine procedures has resulted in a substantial increase in confirmed bacterial meningitis cases. However, in resource-poor countries, these assays are only available in reference laboratories. Sample transportation to these laboratories is a critical constraint, as it requires specialized, high cost courier services. To overcome this barrier we evaluated the use of FTATM Elute filter paper cards for the conservation and processing of samples under normal environmental conditions, as they would be when transported from remote and under-equipped healthcare facilities to the reference centers. A total of 401 samples received in 2015 as part of Sao Paulo's national surveillance for routine diagnosis were selected for this study. METHODS: The sensitivity and specificity of real-time PCR were evaluated using fresh serum and cerebrospinal fluid (CSF) samples processed using our laboratory's standard DNA extraction, and processing the same samples after being dried and stored on FTATM card, and DNA extracted following the manufacturer's instructions. RESULTS: The sensitivities for detection of Neisseria meningitidis, Streptococcus pneumoniae, and Haemophilus influenzae from CSF dried and stored on FTATM cards were 98%, 92%, and 100%, respectively, and with serum samples were 73%, 88%, and 100%, respectively. When compared to our laboratory's standard methodology, results showed high concordance, with Kappa index ranges of 0.9877-1.00 for CSF, and 0.8004-1.00 for serum samples. CONCLUSION: The use of FTATM cards for CSF and serum conservation and transport represents a rapid, reliable, and cost-effective alternative that will allow obtaining valuable epidemiological information that would otherwise be lost.
Subject(s)
Haemophilus influenzae/isolation & purification , Meningitis, Bacterial/blood , Meningitis, Bacterial/cerebrospinal fluid , Neisseria meningitidis/isolation & purification , Streptococcus pneumoniae/isolation & purification , Brazil/epidemiology , Female , Haemophilus influenzae/pathogenicity , Humans , Male , Meningitis, Bacterial/epidemiology , Meningitis, Bacterial/microbiology , Neisseria meningitidis/pathogenicity , Streptococcus pneumoniae/pathogenicityABSTRACT
BACKGROUND: Meningococcal carriage studies are important to improve our understanding of the epidemiology of meningococcal disease. The aim of this study was to determine the prevalence of meningococcal carriage and the phenotypic and genotypic characteristics of isolates collected from a sample of students in the city of Bogotá, Colombia. MATERIALS AND METHODS: A total of 1459 oropharyngeal samples were collected from students aged 15-21 years attending secondary schools and universities. Swabs were plated on a Thayer Martin agar and N. meningitidis was identified by standard microbiology methods and PCR. RESULTS: The overall carriage prevalence was 6.85%. Carriage was associated with cohabitation with smokers, and oral sex practices. Non-groupable and serogroup Y isolates were the most common capsule types found. Isolates presented a high genetic diversity, and circulation of the hypervirulent clonal complexes ST-23, ST-32 and ST-41/44 were detected. CONCLUSION: The meningococcal carriage rate was lower than those reported in Europe and Africa, but higher than in other Latin American countries. Our data also revealed antigenic and genetic diversity of the isolates and the circulation of strains belonging to clonal complexes commonly associated with meningococcal disease.
Subject(s)
Carrier State/microbiology , Meningitis, Meningococcal/microbiology , Neisseria meningitidis/isolation & purification , Adolescent , Adult , Antigens, Bacterial/immunology , Colombia , Female , Genotype , Humans , Male , Neisseria meningitidis/genetics , Oropharynx/microbiology , Prevalence , Serotyping/methods , Students , Young AdultABSTRACT
BACKGROUND: With 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. METHODS: A 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. RESULTS: Serogroup 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. CONCLUSIONS: Our 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.
Subject(s)
Neisseria meningitidis/genetics , Bacterial Typing Techniques , Chile , Humans , Meningococcal Infections/microbiology , Multilocus Sequence Typing , Neisseria meningitidis/classification , Neisseria meningitidis/pathogenicity , SerotypingABSTRACT
Phenotypic and genotypic characterization of 133 isolates of Neisseria meningitidis obtained from meningococcal disease cases in Argentina during 2010 were performed by the National Reference Laboratory as part of a project coordinated by the PAHO within the SIREVA II network. Serogroup, serotype, serosubtype and MLST characterization were performed. Minimum Inhibitory Concentration to penicillin, ampicillin, ceftriaxone, rifampin, chloramphenicol, tetracycline and ciprofloxacin were determined and interpreted according to CLSI guidelines. Almost 49% of isolates were W135, and two serotype:serosubtype combinations, W135:2a:P1.5,2:ST-11 and W135:2a:P1.2:ST-11 accounted for 78% of all W135 isolates. Serogroup B accounted for 42.1% of isolates, and was both phenotypically and genotypically diverse. Serogroup C isolates represented 5.3% of the dataset, and one isolate belonging to the ST-198 complex was non-groupable. Isolates belonged mainly to the ST-11 complex (48%) and to a lesser extent to the ST-865 (18%), ST-32 (9,8%) and the ST-35 complexes (9%). Intermediate resistance to penicillin and ampicillin was detected in 35.4% and 33.1% of isolates respectively. Two W135:2a:P1.5,2:ST-11:ST-11 isolates presented resistance to ciprofloxacin associated with a mutation in the QRDR of gyrA gene Thr91-Ile. These data show serogroup W135 was the first cause of disease in Argentina in 2010, and was strongly associated with the W135:2a:P1.5,2:ST-11 epidemic clone. Serogroup B was the second cause of disease and isolates belonging to this serogroup were phenotypically and genotypically diverse. The presence of isolates with intermediate resistance to penicillin and the presence of fluorquinolone-resistant isolates highlight the necessity and importance of maintaining and strengthening National Surveillance Programs.
Subject(s)
Meningitis, Meningococcal/microbiology , Neisseria meningitidis/genetics , Anti-Infective Agents/pharmacology , Argentina , Ciprofloxacin/pharmacology , Drug Resistance, Bacterial/drug effects , Drug Resistance, Bacterial/genetics , Genotype , Humans , Microbial Sensitivity Tests , Neisseria meningitidis/classification , Neisseria meningitidis/drug effects , Neisseria meningitidis/isolation & purification , PhenotypeABSTRACT
BACKGROUND: Published data on the epidemiology of meningococcal disease in Latin America and the Caribbean region is scarce and, when available, it is often published in Spanish and/or in non-peer-reviewed journals, making it difficult for the international scientific community to have access. METHODS: Laboratory data on 4,735 Neisseria meningitidis strains was collected and reported by the National Reference Laboratories in 19 Latin American countries and the Caribbean Epidemiology Centre (CAREC) between 2006 and 2010 as part of the work carried out by the SIREVA II network. Serogroup and MIC to penicillin, rifampin and chloramphenicol were determined. RESULTS: Isolates were mainly obtained from patients <5 years, but each year around 25% of isolates came from adult patients. Serogroup distribution was highly variable among countries. Serogroup C was the main cause of disease in Brazil; the majority of disease seen in the Southern cone was caused by serogroup B, but serogroup W135 strains have increased in recent years. In the Andean and Mexico, Central America and Caribbean regions, serogroups B and C were equally present, and serogroup Y was frequently isolated. Isolates were generally susceptible to chloramphenicol, penicillin and rifampin, but almost 60% of isolates characterized in Southern cone countries presented intermediate resistance to penicillin. Five rifampin-resistant isolates have been isolated in Uruguay and Brazil. CONCLUSIONS: Serogroup distribution is highly variable among countries, but some geographic structuring can be inferred from these data. Epidemiological and laboratory data are scarce among Andean and Mexico, Central America and Caribbean countries. Evaluation and implementation of corrective measures on disease surveillance and reporting systems and the implementation of molecular diagnostic techniques and molecular characterization on meningococcal isolates are advised.
Subject(s)
Clinical Laboratory Techniques/methods , Meningococcal Infections/epidemiology , Meningococcal Infections/microbiology , Neisseria meningitidis/classification , Population Surveillance/methods , Adolescent , Adult , Age Distribution , Anti-Bacterial Agents/pharmacology , Caribbean Region/epidemiology , Child , Child, Preschool , Humans , Infant , Infant, Newborn , Latin America/epidemiology , Microbial Sensitivity Tests , Neisseria meningitidis/drug effects , Neisseria meningitidis/isolation & purification , Penicillins/pharmacology , Rifampin/pharmacology , Serotyping , Young AdultABSTRACT
BACKGROUND: Herd immunity is important in the effectiveness of conjugate polysaccharide vaccines against encapsulated bacteria. A large multicenter study investigated the effect of meningococcal serogroup C conjugate vaccine introduction on the meningococcal population. METHODS: Carried meningococci in individuals aged 15-19 years attending education establishments were investigated before and for 2 years after vaccine introduction. Isolates were characterized by multilocus sequence typing, serogroup, and capsular region genotype and changes in phenotypes and genotypes assessed. RESULTS: A total of 8462 meningococci were isolated from 47 765 participants (17.7%). Serogroup prevalence was similar over the 3 years, except for decreases of 80% for serogroup C and 40% for serogroup 29E. Clonal complexes were associated with particular serogroups and their relative proportions fluctuated, with 12 statistically significant changes (6 up, 6 down). The reduction of ST-11 complex serogroup C meningococci was probably due to vaccine introduction. Reasons for a decrease in serogroup 29E ST-254 meningococci (from 1.8% to 0.7%) and an increase in serogroup B ST-213 complex meningococci (from 6.7% to 10.6%) were less clear. CONCLUSIONS: Natural fluctuations in carried meningococcal genotypes and phenotypes a can be affected by the use of conjugate vaccines, and not all of these changes are anticipatable in advance of vaccine introduction.
Subject(s)
Immunity, Herd/immunology , Meningitis, Meningococcal/prevention & control , Meningococcal Vaccines/administration & dosage , N-Acetylneuraminic Acid/genetics , Neisseria meningitidis/genetics , Neisseria meningitidis/immunology , Adolescent , Adult , Bacterial Capsules/genetics , Bacterial Capsules/metabolism , Carrier State/immunology , Genotype , Humans , Mass Vaccination , Meningitis, Meningococcal/genetics , Meningitis, Meningococcal/immunology , Multilocus Sequence Typing , N-Acetylneuraminic Acid/metabolism , Serotyping , United Kingdom , Young AdultABSTRACT
BACKGROUND: The epidemiology of meningococcal disease in Mozambique and other African countries located outside the "meningitis belt" remains widely unknown. With the event of upcoming vaccines microbiological and epidemiological information is urgently needed. METHODS: Prospective surveillance for invasive bacterial infections was conducted at the Manhiça District hospital (rural Mozambique) among hospitalized children below 15 years of age. Available Neisseria meningitidis isolates were serogrouped and characterized by Multilocus Sequence Typing (MLST). Antibiotic resistance was also determined. RESULTS: Between 1998 and 2008, sixty-three cases of confirmed meningococcal disease (36 meningitis, 26 sepsis and 1 conjunctivitis) were identified among hospitalized children. The average incidence rate of meningococcal disease was 11.6/100,000 (8/100,000 for meningitis and 3.7/100,000 for meningococcemia, respectively). There was a significant rise on the number of meningococcal disease cases in 2005-2006 that was sustained till the end of the surveillance period. Serogroup was determined for 43 of the 63 meningococcal disease cases: 38 serogroup W-135, 3 serogroup A and 2 serogroup Y. ST-11 was the most predominant sequence type and strongly associated with serogroup W-135. Two of the three serogroup A isolates were ST-1, and both serogroup Y isolates were ST-175. N. meningitidis remained highly susceptible to all antibiotics used for treatment in the country, although the presence of isolates presenting intermediate resistance to penicillin advocates for continued surveillance. CONCLUSIONS: Our data show a high rate of meningococcal disease in Manhiça, Mozambique, mainly caused by serogroup W-135 ST-11 strains, and advocates for the implementation of a vaccination strategy covering serogroup W-135 meningococci in the country.
Subject(s)
Drug Resistance, Bacterial , Meningococcal Infections/epidemiology , Neisseria meningitidis/physiology , Rural Population/statistics & numerical data , Adolescent , Bacteremia , Body Fluids/microbiology , Child , Child, Preschool , Humans , Incidence , Infant , Meningococcal Infections/microbiology , Microbial Sensitivity Tests , Mozambique/epidemiology , Neisseria meningitidis/classification , Neisseria meningitidis/genetics , Neisseria meningitidis/isolation & purification , SerotypingABSTRACT
The epidemiology of meningococcal disease in Mozambique and other African countries located outside the "meningitis belt" remains widely unknown. With the event of upcoming vaccines microbiological and epidemiological information is urgently needed. Prospective surveillance for invasive bacterial infections was conducted at the Manhiça District hospital (rural Mozambique) among hospitalized children below 15 years of age. Available Neisseria meningitidis isolates were serogrouped and characterized by Multilocus Sequence Typing (MLST). Antibiotic resistance was also determined. Between 1998 and 2008, sixty-three cases of confirmed meningococcal disease (36 meningitis, 26 sepsis and 1 conjunctivitis) were identified among hospitalized children. The average incidence rate of meningococcal disease was 11.6/100,000 (8/100,000 for meningitis and 3.7/100,000 for meningococcemia, respectively). There was a significant rise on the number of meningococcal disease cases in 20052006 that was sustained till the end of the surveillance period. Serogroup was determined for 43 of the 63 meningococcal disease cases: 38 serogroup W-135, 3 serogroup A and 2 serogroup Y. ST-11 was the most predominant sequence type and strongly associated with serogroup W-135. Two of the three serogroup A isolates were ST-1, and both serogroup Y isolates were ST175. N.meningitidis remained highly susceptible to all antibiotics used for treatment in the country, although the presence of isolates presenting intermediate resistance to penicillin advocates for continued surveillance.Our data show a high rate of meningococcal disease in Manhiça, Mozambique, mainly caused by serogroup W-135 ST-11 strains, and advocates for the implementation of a vaccination strategy covering serogroup W-135 meningococci in the country.
Subject(s)
Humans , Meningitis, Bacterial/immunology , Drug Resistance, Bacterial , Meningococcal Infections/epidemiology , Neisseria meningitidis/physiology , Rural Population/statistics & numerical data , MozambiqueABSTRACT
Previously we have shown that insertion of IS1301 in the sia/ctr intergenic region (IGR) of serogroup C Neisseria meningitidis (MenC) isolates from Spain confers increased resistance against complement-mediated killing. Here we investigate the significance of IS1301 in the same location in N. meningitidis isolates from the UK. PCR and sequencing was used to screen a collection of more than 1500 meningococcal carriage and disease isolates from the UK for the presence of IS1301 in the IGR. IS1301 was not identified in the IGR among vaccine failure strains but was frequently found in serogroup B isolates (MenB) from clonal complex 269 (cc269). Almost all IS1301 insertions in cc269 were associated with novel polymorphisms, and did not change capsule expression or resistance to human complement. After excluding sequence types (STs) distant from the central genotype within cc269, there was no significant difference for the presence of IS1301 in the IGR of carriage isolates compared to disease isolates. Isolates with insertion of IS1301 in the IGR are not responsible for MenC disease in UK vaccine failures. Novel polymorphisms associated with IS1301 in the IGR of UK MenB isolates do not lead to the resistance phenotype seen for IS1301 in the IGR of MenC isolates.
Subject(s)
Bacterial Capsules/biosynthesis , DNA Transposable Elements/genetics , Meningococcal Infections/immunology , Neisseria meningitidis, Serogroup C/immunology , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Base Sequence , Complement System Proteins/immunology , DNA, Bacterial/genetics , DNA, Intergenic/genetics , Humans , Meningococcal Infections/microbiology , Meningococcal Infections/prevention & control , Meningococcal Vaccines/administration & dosage , Meningococcal Vaccines/immunology , Mutagenesis, Insertional , Neisseria meningitidis, Serogroup B/classification , Neisseria meningitidis, Serogroup B/genetics , Neisseria meningitidis, Serogroup B/immunology , Neisseria meningitidis, Serogroup C/classification , Neisseria meningitidis, Serogroup C/genetics , Phylogeny , Sequence Homology, Nucleic Acid , Spain , United KingdomABSTRACT
Multilocus sequence typing (MLST) was first proposed in 1998 as a typing approach that enables the unambiguous characterization of bacterial isolates in a standardized, reproducible, and portable manner using the human pathogen Neisseria meningitidis as the exemplar organism. Since then, the approach has been applied to a large and growing number of organisms by public health laboratories and research institutions. MLST data, shared by investigators over the world via the Internet, have been successfully exploited in applications ranging from molecular epidemiological investigations to population biology and evolutionary analyses. This chapter describes the practical steps in the development and application of an MLST scheme and some of the common tools and techniques used to obtain the maximum benefit from the data. Considerations pertinent to the implementation of high-capacity MLST projects (i.e., those involving thousands of isolates) are discussed.
Subject(s)
Bacteria/classification , Bacteria/genetics , Bacterial Typing Techniques/methods , Molecular Epidemiology/methods , Bacteria/isolation & purification , DNA, Bacterial/genetics , DNA, Bacterial/isolation & purification , Databases, Nucleic Acid , Electrophoresis, Agar Gel , Evolution, Molecular , Genetics, Population , Humans , Models, Genetic , Neisseria meningitidis/classification , Neisseria meningitidis/genetics , Neisseria meningitidis/isolation & purification , Nucleic Acid Amplification Techniques , Polymerase Chain Reaction , Sequence Analysis, DNAABSTRACT
The meningococcal Opa proteins play an important role in pathogenesis by mediating invasion of human cells. The aim of this investigation was to determine whether carried and disease-associated meningococci possess different Opa repertoires and whether the diversity of these proteins is associated with clinical severity of disease. Opa repertoires in 227 disease-associated meningococci, isolated in the United Kingdom over a period of 6 years, were compared to the repertoires in 190 asymptomatically carried meningococci isolated in the United Kingdom from a contemporary, nonepidemic period. Multidimensional scaling (MDS) was employed to investigate the association between Opa repertoires and multilocus sequence typing (MLST) genotypes. Associations with clinical severity were also analyzed statistically. High levels of diversity were observed in opa alleles, variable regions, and repertoires, and MDS revealed that MLST genotypes were strongly associated with particular Opa repertoires. Individual Opa proteins or repertoires were not associated with clinical severity, though there was a trend toward an association with the opaD locus. Meningococcal Opa repertoire is strongly linked to MLST genotype irrespective of epidemiological sampling and therefore correlates with invasiveness. It is not, however, strongly associated with severity of meningococcal disease.
Subject(s)
Bacterial Outer Membrane Proteins/genetics , Adolescent , DNA, Bacterial/genetics , Genetic Variation , Humans , Meningococcal Infections/microbiology , Molecular Sequence Data , Neisseria meningitidis/genetics , Neisseria meningitidis/pathogenicity , Polymerase Chain Reaction , Young AdultABSTRACT
BACKGROUND: In 1999, meningococcal serogroup C conjugate (MCC) vaccines were introduced in the United Kingdom for those under 19 years of age. The impact of this intervention on asymptomatic carriage of meningococci was investigated to establish whether serogroup replacement or protection by herd immunity occurred. METHODS: Multicenter surveys of carriage were conducted during vaccine introduction and on 2 successive years, resulting in a total of 48,309 samples, from which 8599 meningococci were isolated and characterized by genotyping and phenotyping. RESULTS: A reduction in serogroup C carriage (rate ratio, 0.19) was observed that lasted at least 2 years with no evidence of serogroup replacement. Vaccine efficacy against carriage was 75%, and vaccination had a disproportionate impact on the carriage of sequence type (ST)-11 complex serogroup C meningococci that (rate ratio, 0.06); these meningococci also exhibited high rates of capsule expression. CONCLUSIONS: The impact of vaccination with MCC vaccine on the prevalence of carriage of group C meningococci was consistent with herd immunity. The high impact on the carriage of ST-11 complex serogroup C could be attributed to high levels of capsule expression. High vaccine efficacy against disease in young children, who were not protected long-term by the schedule initially used, is attributed to the high vaccine efficacy against carriage in older age groups.