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An effective contagious caprine pleuropneumonia (CCPP) vaccine is essential for the increased production of healthy goats in a cost-effective manner and the prevention of animal-to-animal transmission for both domestic animals and wildlife. Quality control of this vaccine ensures that a reliable supply of pure, safe, and potent batches is obtained. As part of this control, in vitro quantification of Mycoplasma capricolum subsp. capripneumoniae (Mccp) protein in the final vaccines is required before the CCPP vaccine undergoes batch release and certification. The current method used for quantification is based on the measurement of total protein using the bicinchonic acid (BCA) test. This method quantifies the total amount of protein in the vaccine including contaminant protein from media, which can lead to overestimation of the quantity of Mccp protein, resulting in reduced vaccine immunogenicity. An immuno-capture ELISA (ICE) was developed for specific detection and quantification of the Mccp antigen in the CCPP vaccine. As the ICE detects and measures the amount of antigen between two layers of antibodies, capture and detecting antibodies are required. Mouse monoclonal antibodies (mAbs) that detect the Mccp antigen were produced and characterized. One of these mAbs, Mccp-25, was used to develop the ICE as an unlabelled capture antibody and horseradish peroxidase conjugated detecting antibody. The ICE was standardized and evaluated using an internal reference sample, experimental CCPP vaccines and commercial CCPP vaccines. A comparison between the polymerase chain reaction (PCR) and ICE showed good correlation between the two assays. Also, an in vitro ICE method correlated well with an in vivo sero-conversion in goats that were vaccinated with selected test vaccines. The sensitivity of the ICE was estimated at 30 ng/ml.

Enzyme linked immunosorbent assays (ELISAs) have been developed for the detection of antibodies against contagious caprine pleuropneumonia (CCPP), the causative agent of which is Mycoplasma capricolum subsp. Capripneumoniae (Mccp). The currently available commercial CCPP competitive ELISA (CCPP cELISA) kit produced and supplied by IDEXX Company (Westbrook, Maine, United States) is relatively expensive for most African laboratories. To address this issue and provide a variety of choices, a sensitive and specific blocking-ELISA (b-ELISA) test to detect antibodies against CCPP was developed. We describe the newly developed CCPP blocking-ELISA based on the blocking of an epitope of a monoclonal antibody (Mccp-25) by a positive serum sample against the Mccp protein coated on a plate. The Percentage Inhibition (PI) cut-off value for the CCPP b-ELISA was set at 50 using 466 CCPP negative and 84 CCPP positive small ruminant sera. Of the negative sera, 307 were obtained from the Botswana National Veterinary Laboratory (BNVL) and 159 from the Friedrich-Loeffler-Institute (FLI) Germany. The 84 positive sera samples came from experimentally vaccinated goats at the AU-PANVAC facility in Debre-Zeit, Ethiopia. The relative diagnostic sensitivity and specificity of the CCPP b-ELISA was 93% and 88%, respectively. This test result indicated good correlation with that of the commercial CCPP cELISA by IDEXX Company (Westbrook, Maine, United States) with a Cohen’s κ agreement of κ agreement of 0.85. The newly developed CCPP b-ELISA will be useful in the detection of antibodies for the diagnosis CCPP and for sero-surveillance during vaccination campaigns.

Efforts to prevent the large-scale outbreaks of meningococcal disease that have led to significant morbidity and mortality in the African meningitis belt, a region stretching from Senegal to Ethiopia, changed dramatically in 2010 when a newly developed group A meningococcal polysaccharide–tetanus toxoid protein conjugate vaccine, PsA-TT or MenAfriVac, was introduced in the first-ever preventive mass vaccination campaign in Burkina Faso, Mali, and Niger [1–4]. Developed by the Meningitis Vaccine Project, a partnership between the World Health Organization (WHO) and PATH with funding from the Bill & Melinda Gates Foundation, PsA-TT transformed the reactive vaccination strategy in place in the Meningitis Belt since the 1970s, which utilized polysaccharide meningococcal vaccines to respond to meningococcal disease outbreaks, into a preventive campaign that is implementing mass vaccination of individuals aged 1–29 years in >25 countries in Africa [5]. Evidence has indicated that PsA-TT is highly immunogenic; the vaccine was licensed based on immunogenicity alone [2, 6–8]. By the end of 2014, >217 million people in 15 countries had been vaccinated with PsA-TT [9]. The introduction of PsA-TT has dramatically altered the epidemiology of meningococcal disease in Africa, and significant reductions in the incidence of meningococcal A disease [10, 11] and meningococcal A carriage [12, 13] have been achieved in countries where the vaccine has been introduced. As with all recently developed and newly introduced vaccines, questions remain about the duration of protection provided by PsA-TT. Assessing the duration of protection has been identified as a priority for the prevention and control of meningitis in Africa [14]. Evaluating antibody persistence both in the context of clinical trials and in population-based epidemiologic studies is an important component of investigating the impact conjugate meningococcal A vaccination has had on immunity in this region. Meningococcal conjugate vaccines including PsA-TT join together polysaccharides from the outer membrane of Neisseria meningitidis to a protein carrier such as tetanus toxoid. Conjugate vaccines elicit a more robust immune response than polysaccharide-only vaccines, which are limited in that immunity wanes after 3–5 years and in that they are not immunogenic in very young children [15–17]. Although conjugate meningococcal C and ACWY vaccines provide a longer duration of immunity than comparable polysaccharide vaccines, antibody levels have been shown to wane rapidly following vaccination [18, 19]. Booster doses have been added to vaccination schedules in both the United States, where meningococcal ACWY conjugate vaccine is recommended for 11- and 12-year-olds with a booster dose at age 16 years [20], and the United Kingdom, where meningococcal C conjugate vaccine is recommended for infants and booster doses are given at 12 months and around 14 years [21]. Booster doses for these meningococcal conjugate vaccines have been recommended due to concerns about waning immunity during periods of high risk [18, 22–24], further emphasizing the need to understand the duration of immunity following PsA-TT introduction has been introduced in areas where meningococcal disease is highly endemic. To assess the duration of protection and changes in population-level immunity over time following the 2010 introduction of PsA-TT, we established a cohort in 2012 among residents of Bamako, Mali. Here we report the results of the first seroprevalence survey undertaken in the cohort, 2 years after the PsA-TT mass vaccination campaign.

The pattern of epidemic meningococcal disease in the African meningitis belt may be influenced by the background level of population immunity but this has been measured infrequently. A standardised enzyme-linked immunosorbent assay (ELISA) for measuring meningococcal serogroup A IgG antibodies was established at five centres within the meningitis belt. Antibody concentrations were then measured in 3930 individuals stratified by age and residence from six countries. Seroprevalence by age was used in a catalytic model to determine the force of infection. Meningococcal serogroup A IgG antibody concentrations were high in each country but showed heterogeneity across the meningitis belt. The geometric mean concentration (GMC) was highest in Ghana (9.09 μg/mL [95% CI 8.29, 9.97]) and lowest in Ethiopia (1.43 μg/mL [95% CI 1.31, 1.57]) on the margins of the belt. The force of infection was lowest in Ethiopia (λ = 0.028). Variables associated with a concentration above the putative protective level of 2 μg/mL were age, urban residence and a history of recent vaccination with a meningococcal vaccine. Prior to vaccination with the serogroup A meningococcal conjugate vaccine, meningococcal serogroup A IgG antibody concentrations were high across the African meningitis belt and yet the region remained susceptible to epidemics.

Summary Background Information on transmission of meningococcal infection in the African meningitis belt is scarce. We aimed to describe transmission patterns of Neisseria meningitidis (meningococcus) in households in the African meningitis belt. Methods Cross-sectional carriage surveys were done in seven African meningitis belt countries (Chad, Ethiopia, Ghana, Mali, Niger, Nigeria, and Senegal) between Aug 1, 2010, and Oct 15, 2012. Meningococcal carriers identified in these surveys and all available people in their households were recruited into this longitudinal cohort study. We took pharyngeal swabs at first visit and took further swabs twice a month for 2 months and then monthly for a further 4 months. We used conventional bacteriological and molecular techniques to identify and characterise meningococci. We estimated the rates of carriage acquisition and recovery using a multi-state Markov model. Findings Meningococci were isolated from 241 (25%) of 980 members of 133 households in which a carrier had been identified in the cross-sectional survey or at the first household visit. Carriage was detected subsequently in another household member who was not an index carrier in 75 households. Transmission within a household, suggested by detection of a further carrier with the same strain as the index carrier, was found in 52 of these 75 households. Children younger than 5 years were the group that most frequently acquired carriage from other household members. The overall individual acquisition rate was 2·4% (95% CI 1·6–4·0) per month, varying by age and household carriage status. The mean duration of carriage was 3·4 months (95% CI 2·7–4·4). Interpretation In the African meningitis belt, transmission of meningococci within households is important, particularly for young children, and periods of carriage are usually of short duration. Funding Bill & Melinda Gates Foundation, Wellcome Trust.

To investigate potential risk factors for acquisition in seven countries of the meningitis belt.Households were followed up every 2 weeks for 2 months, then monthly for a further 4 months. Pharyngeal swabs were collected from all available household members at each visit and questionnaires completed. Risks of acquisition over the whole study period and for each visit were analysed by a series of logistic regressions.Over the course of the study, acquisition was higher in: (i) 5-to 14-year olds, as compared with those 30 years or older (OR 3.6, 95% CI 1.4-9.9); (ii) smokers (OR 3.6, 95% CI 0.98-13); and (iii) those exposed to wood smoke at home (OR 2.6 95% CI 1.3-5.6). The risk of acquisition from one visit to the next was higher in those reporting a sore throat during the dry season (OR 3.7, 95% CI 2.0-6.7) and lower in those reporting antibiotic use (OR 0.17, 95% CI 0.03-0.56).Acquisition of meningococcal carriage peaked in school age children. Recent symptoms of sore throat during the dry season, but not during the rainy season, were associated with a higher risk of acquisition. Upper respiratory tract infections may be an important driver of epidemics in the meningitis belt.Investiguer les facteurs de risque potentiels d'acquisition dans sept pays de la ceinture de la méningite. MÉTHODES: Des ménages ont été suivis toutes les deux semaines pendant deux mois, puis tous les mois pendant quatre mois. Des prélèvements pharyngés sur écouvillons ont été collectés auprès de tous les membres disponibles du ménage à chaque visite et des questionnaires ont été remplis. Les risques d'acquisition sur l'ensemble de la période d’étude et pour chaque visite ont été analysés par une série de régressions logistiques. RÉSULTATS: Au cours de l’étude, l'acquisition a été plus élevée chez: (i) les 5-14 ans, par rapport à ceux âgés de 30 ans ou plus (OR = 3,6; IC95%: 1,4-9,9); (ii) les fumeurs (OR = 3,6; IC95%: 0,98-13); et (iii) les personnes exposées à la fumée de bois à la maison (OR = 2,6; IC95%: 1,3-5,6). Le risque d'acquisition d'une visite à l'autre était plus élevé chez les personnes signalant un mal de gorge pendant la saison sèche (OR = 3,7; IC95%: 2,0-6,7) et plus faible chez celles signalant une utilisation d'antibiotique (OR = 0,17; IC95%: 0,03-0,56).L'acquisition du portage du méningocoque a culminé chez les enfants d’âge scolaire. Les symptômes récents de maux de gorge pendant la saison sèche, mais pas pendant la saison des pluies, étaient associés à un risque d'acquisition plus élevé. Les infections des voies respiratoires supérieures pourraient être un facteur important d’épidémies dans la ceinture de la méningite.

International audience; Background. Study of meningococcal carriage is essential to understanding the epidemiology of Neisseria meningitidis infection.Methods. Twenty cross-sectional carriage surveys were conducted in 7 countries in the African meningitis belt; 5 surveys were conducted after introduction of a new serogroup A meningococcal conjugate vaccine (MenAfriVac). Pharyngeal swab specimens were collected, and Neisseria species were identified by microbiological and molecular techniques.Results. A total of 1687 of 48 490 participants (3.4%; 95% confidence interval [CI], 3.2%–3.6%) carried meningococci. Carriage was more frequent in individuals aged 5–14 years, relative to those aged 15–29 years (adjusted odds ratio [OR], 1.41; 95% CI, 1.25–1.60); in males, relative to females (adjusted OR, 1.17; 95% CI, 1.10–1.24); in individuals in rural areas, relative to those in urban areas (adjusted OR, 1.44; 95% CI, 1.28–1.63); and in the dry season, relative to the rainy season (adjusted OR, 1.54; 95% CI, 1.37–1.75). Forty-eight percent of isolates had genes encoding disease-associated polysaccharide capsules; genogroup W predominated, and genogroup A was rare. Strain diversity was lower in countries in the center of the meningitis belt than in Senegal or Ethiopia. The prevalence of genogroup A fell from 0.7% to 0.02% in Chad following mass vaccination with MenAfriVac.Conclusions. The prevalence of meningococcal carriage in the African meningitis belt is lower than in industrialized countries and is very diverse and dynamic, even in the absence of vaccination.

A meningococcal serogroup A polysaccharide/tetanus toxoid conjugate vaccine (PsA-TT) (MenAfriVac#x2122;) is being deployed in countries of the African meningitis belt. Experience with other polysaccharide/protein conjugate vaccines has shown that an important part of their success has been their ability to prevent the acquisition of pharyngeal carriage and hence to stop transmission and induce herd immunity. If PsA-TT is to achieve the goal of preventing epidemics, it must be able to prevent the acquisition of pharyngeal carriage as well as invasive meningococcal disease and whether PsA-TT can prevent pharyngeal carriage needs to be determined. To address this issue, a consortium (the African Meningococcal Carriage (MenAfriCar) consortium) was established in 2009 to investigate the pattern of meningococcal carriage in countries of the African meningitis belt prior to and after the introduction of PsA-TT. This article describes how the consortium was established, its objectives and the standardised field and laboratory methods that were used to achieve these objectives. The experience of the MenAfriCar consortium will help in planning future studies on the epidemiology of meningococcal carriage in countries of the African meningitis belt and elsewhere. Un vaccin conjugué contenant un polysaccharide du sérogroupe A méningococcique et une anatoxine du tétanos (PsA-TT) (MenAfriVac™) est en cours de déploiement dans les pays de la ceinture africaine de la méningite. L’ expérience avec d’ autres vaccins conjugués polysaccharide/protéine a montré qu’ une partie importante de leur succès a été leur capacité à empêcher l’ acquisition du portage pharyngé et donc à arrêter la transmission et à induire une immunité de group. Si PsA-TT doit d’ atteindre l’ objectif de prévenir les épidémies, il devrait être en mesure d’ empêcher l’ acquisition du portage pharyngé ainsi que la méningococcie invasive et le fait que PsA-TT puisse empêcher le portage pharyngé devrait être déterminé. Pour résoudre ce problème, le consortium MenAfriCar (Consortium Africain du Portage Méningococcique) a été établi en 2009 pour étudier le mode de portage du méningocoque dans les pays de la ceinture africaine de la méningite avant et après l’ introduction de PsA-TT. Cet article décrit comment le consortium a été établi, ses objectifs et les méthodes de laboratoire et de terrain standardisées qui ont été utilisées pour atteindre ces objectifs. L’ expérience du consortium MenAfriCar aidera à planifier les futures études sur l’ épidémiologie du portage du méningocoque dans les pays de la ceinture africaine de la méningite et d’ ailleurs. Se está utilizando una vacuna meningocócica conjugada (MenAfriVac™) de polisacárido del serogrupo A / tétano toxoide (PsA-TT) en países del cinturón Africano de meningitis. Las experiencias obtenidas con otras vacunas conjugadas polisacárido/proteína han demostrado que una parte importante de su éxito se debe a su habilidad para prevenir la colonización faríngea de los portadores, acabando por lo tanto con la transmisión, y a la de inducir la protección de rebaño. Si PsA-TT ha de cumplir el objetivo de prevenir epidemias, debe ser capaz de prevenir el estado de portador faríngeo, al igual que la enfermedad invasiva por meningococo, y para ello es necesario determinar si la PsA-TT puede prevenir la colonización faríngea. Con el fin de abordar esta cuestión se estableció un consorcio africano en el 2009 - el MenAfriCar (African Meningococcal Carriage Consortium) – para investigar los patrones del estado de portador de meningococo en países del cinturón Africano de la meningitis, antes y después de la introducción de PsA-TT. Este artículo describe como se estableció el consorcio, sus objetivos y los métodos estandarizados de campo y de laboratorio que se utilizaron para alcanzarlos. La experiencia del consorcio MenAfriCar ayudará en la planificación de estudios futuros sobre la epidemiología del estado de portador de meningococo, tanto en países del cinturón Africano de la meningitis como en otros lugares del mundo.