Modeling Optimal Laboratory Testing Strategies for Bacterial Meningitis Surveillance in Africa.

Since 2010, the introduction of an effective serogroup A meningococcal conjugate vaccine has led to the near-elimination of invasive Neisseria meningitidis serogroup A disease in Africa's meningitis belt. However, a significant burden of disease and epidemics due to other bacterial meningitis pathogens remain in the region. High-quality surveillance data with laboratory confirmation is important to monitor circulating bacterial meningitis pathogens and design appropriate interventions, but complete testing of all reported cases is often infeasible. Here, we use case-based surveillance data from 5 countries in the meningitis belt to determine how accurately estimates of the distribution of causative pathogens would represent the true distribution under different laboratory testing strategies. Detailed case-based surveillance data was collected by the MenAfriNet surveillance consortium in up to 3 seasons from participating districts in 5 countries. For each unique country-season pair, we simulated the accuracy of laboratory surveillance by repeatedly drawing subsets of tested cases and calculating the margin of error of the estimated proportion of cases caused by each pathogen (the greatest pathogen-specific absolute error in proportions between the subset and the full set of cases). Across the 12 country-season pairs analyzed, the 95% credible intervals around estimates of the proportion of cases caused by each pathogen had median widths of ±0.13, ±0.07, and ±0.05, respectively, when random samples of 25%, 50%, and 75% of cases were selected for testing. The level of geographic stratification in the sampling process did not meaningfully affect accuracy estimates. These findings can inform testing thresholds for laboratory surveillance programs in the meningitis belt.

Personal protective equipment wearing in the context of COVID-19 in a hospital in Burkina Faso / Port des équipements de protection individuelle dans le contexte de la COVID-19 dans un centre hospitalier du Burkina Faso

INTRODUCTION: Personal protective equipment (PPE) is used by health care workers to protect themselves and patients from various exposures such as infectious agents. However, the wearing of this equipment is not always optimal, especially in an epidemic context of COVID-19 transmission. AIM OF THE STUDY: The aim of this study, in the specific context of COVID-19, is to contribute to the improvement of PPE wearing practices by health workers. METHODS: This is a descriptive cross-sectional study conducted in 2020 at the Charles De Gaulle Pediatric University Hospital in Burkina Faso. All health workers in the care units and the laboratory were included. Data were collected using an observation grid during the first situation indicating the wearing of PPE. The assessment of the indications for wearing PPE was based on the recommendations of the French Society of Hospital Hygiene and on the technical guide for the prevention and control of healthcare-associated infections in Burkina Faso. RESULTS: Out of 350 targeted agents, 296 were observed. Gowns, masks and gloves were worn in 95.60%, 96.58% and 97.63% of cases respectively. However, PPE such as goggles (1.56%), apron (11.54%), and tunic and pants (46.28%) were poorly used during medical care. CONCLUSION: The practices of health workers with regard to the wearing of certain PPE are still insufficient. A training and awareness program on PPE should be considered to improve patient and staff safety.

Introduction: Les équipements de protection individuelle (EPI) sont utilisés par le personnel de santé dans le cadre des soins pour se protéger et protéger les patients contre les expositions diverses telles que les agents infectieux. Toutefois, le port de ces équipements n'est pas toujours optimal, surtout dans un contexte épidémique de transmission de la COVID-19. But de l'étude: La présente étude vise, dans le contexte spécifique de la COVID-19, à contribuer à l'amélioration des pratiques de port des EPI des agents de santé. Méthodes: Il s'agit d'une étude transversale descriptive conduite en 2020 au centre hospitalier universitaire pédiatrique Charles-de-Gaulle du Burkina Faso. Tous les agents de santé des unités de soins et du laboratoire ont été inclus. Les données ont été recueillies au moyen d'une grille d'observation lors de la première situation indiquant le port d'un EPI. L'appréciation des indications de port des EPI s'est basée sur les recommandations de la Société française d'hygiène hospitalière et sur le Guide technique de prévention et contrôle des infections associées aux soins au Burkina Faso. Résultats: Sur 350 agents ciblés, 296 ont été observés. Le port de la blouse, du masque et des gants était observé respectivement dans 95,60 %, 96,58 % et 97,63 % des cas. Cependant, les EPI tels que les lunettes de protection (1,56 %), le tablier (11,54 %) et la tenue professionnelle composée d'une tunique et d'un pantalon (46,28 %) étaient faiblement utilisés lors des soins médicaux. Conclusion: Les pratiques des agents de santé vis-à-vis du port de certains EPI restent insuffisantes. Un programme de formation et de sensibilisation sur les EPI doit être envisagé afin de renforcer la sécurité des patients et du personnel.

Identification of Streptococcus suis Meningitis by Direct Triplex Real-Time PCR, Burkina Faso.

Meningitis confirmation in Burkina Faso uses PCR for detecting Streptococcus pneumoniae, Neisseria meningitidis, or Hemophilus influenzae. We identified 38 cases of meningitis among 590 that were PCR-positive for 3 nonpneumococcal streptococcal pathogens, including 21 cases of Streptococcus suis. Among the country's 13 regions, 10 had S. suis-positive cases.

Full Molecular Typing of Neisseria meningitidis Directly from Clinical Specimens for Outbreak Investigation.

Neisseria meningitidis is a leading cause of bacterial meningitis and sepsis worldwide and an occasional cause of meningococcal urethritis. When isolates are unavailable for surveillance or outbreak investigations, molecular characterization of pathogens needs to be performed directly from clinical specimens, such as cerebrospinal fluid (CSF), blood, or urine. However, genome sequencing of specimens is challenging because of low bacterial and high human DNA abundances. We developed selective whole-genome amplification (SWGA), an isothermal multiple-displacement amplification-based method, to efficiently enrich, sequence, and de novo assemble N. meningitidis DNA from clinical specimens with low bacterial loads. SWGA was validated with 12 CSF specimens from invasive meningococcal disease cases and 12 urine specimens from meningococcal urethritis cases. SWGA increased the mean proportion of N. meningitidis reads by 2 to 3 orders of magnitude, enabling identification of at least 90% of the 1,605 N. meningitidis core genome loci for 50% of the specimens. The validated method was used to investigate two meningitis outbreaks recently reported in Togo and Burkina Faso. Twenty-seven specimens with low bacterial loads were processed by SWGA before sequencing, and 12 of 27 were successfully assembled to obtain the full molecular typing and vaccine antigen profile of the N. meningitidis pathogen, thus enabling thorough characterization of outbreaks. This method is particularly important for enhancing molecular surveillance in regions with low culture rates. SWGA produces enough reads for phylogenetic and allelic analysis at a low cost. More importantly, the procedure can be extended to enrich other important human bacterial pathogens.

Impact of 13-Valent Pneumococcal Conjugate Vaccine on Pneumococcal Meningitis, Burkina Faso, 2016-2017.

BACKGROUND: In 2013, Burkina Faso introduced 13-valent pneumococcal conjugate vaccine (PCV13) into the routine childhood immunization program, to be administered to children at 8, 12, and 16 weeks of age. We evaluated the impact of PCV13 on pneumococcal meningitis. METHODS: Using nationwide surveillance, we gathered demographic/clinical information and cerebrospinal fluid (CSF) results for meningitis cases. Pneumococcal cases were confirmed by culture, polymerase chain reaction (PCR), or latex agglutination; strains were serotyped using PCR. We compared annual incidence (cases per 100 000) 4 years after PCV13's introduction (2017) to average pre-PCV13 incidence (2011-2013). We adjusted incidence for age and proportion of cases with CSF tested at national laboratories. RESULTS: In 2017, pneumococcal meningitis incidence was 2.7 overall and 10.5 (<1 year), 3.8 (1-4 years), 3.5 (5-14 years), and 1.4 (≥15 years) by age group. Compared to 2011-2013, PCV13-serotype incidence was significantly lower among all age groups, with the greatest decline among children aged <1 year (77%; 95% confidence interval [CI], 65%-84%). Among all ages, the drop in incidence was larger for PCV13 serotypes excluding serotype 1 (79%; 95% CI, 72%-84%) than for serotype 1 (52%; 95% CI, 44%-59%); incidence of non-PCV13 serotypes also declined (53%; 95% CI, 37%-65%). In 2017, 45% of serotyped cases among all ages were serotype 1 and 12% were other PCV13 serotypes. CONCLUSIONS: In Burkina Faso, meningitis caused by PCV13 serotypes continues to decrease, especially among young children. However, the concurrent decline in non-PCV13 serotypes and short pre-PCV13 observation period complicate evaluation of PCV13's impact. Efforts to improve control of serotype 1, such as switching from a 3 + 0 schedule to a 2 + 1 schedule, may improve overall control of pneumococcal meningitis in this setting.

Improving Case-Based Meningitis Surveillance in 5 Countries in the Meningitis Belt of Sub-Saharan Africa, 2015-2017.

BACKGROUND: The MenAfriNet consortium was established in 2014 to support implementation of case-based meningitis surveillance in 5 countries in the meningitis belt of sub-Saharan Africa: Burkina Faso, Chad, Mali, Niger, and Togo. Assessing surveillance performance is critical for interpretation of the collected data and implementation of future surveillance-strengthening initiatives. METHODS: Detailed epidemiologic and laboratory data were collected on suspected meningitis cases through case-based meningitis surveillance in participating districts in 5 countries. Performance of case-based surveillance was evaluated through sensitivity of case ascertainment in case-based versus aggregate meningitis surveillance and an analysis of surveillance indicators. RESULTS: From 2015 to 2017, 18 262 suspected meningitis cases were identified through case-based surveillance and 16 262 were identified through aggregate surveillance, for a case ascertainment sensitivity of 112.3%. Among suspected cases, 16 885 (92.5%) had a cerebrospinal fluid (CSF) specimen collected, 13 625 (80.7%) of which were received at a national reference laboratory. Among these, 13 439 (98.6%) underwent confirmatory testing, and, of those tested, 4371 (32.5%) were confirmed for a bacterial pathogen. CONCLUSIONS: Overall strong performance for case ascertainment, CSF collection, and laboratory confirmation provide evidence for the quality of MenAfriNet case-based surveillance in evaluating epidemiologic trends and informing future vaccination strategies.

Neisseria meningitidis serogroup W, Burkina Faso, 2012.

In 2010, Burkina Faso became the first country to introduce meningococcal serogroup A conjugate vaccine (PsA-TT). During 2012, Burkina Faso reported increases in Neisseria meningitidis serogroup W, raising questions about whether these cases were a natural increase in disease or resulted from serogroup replacement after PsA-TT introduction. We analyzed national surveillance data to describe the epidemiology of serogroup W and genotyped 61 serogroup W isolates. In 2012, a total of 5,807 meningitis cases were reported through enhanced surveillance, of which 2,353 (41%) were laboratory confirmed. The predominant organism identified was N. meningitidis serogroup W (62%), and all serogroup W isolates characterized belonged to clonal complex 11. Although additional years of data are needed before we can understand the epidemiology of serogroup W after PsA-TT introduction, these data suggest that serogroup W will remain a major cause of sporadic disease and has epidemic potential, underscoring the need to maintain high-quality case-based meningitis surveillance after PsA-TT introduction.

Characterization of Plasmodium Falciparum Resistance Genes to Common Antimalarial Drugs in Semi-urban Areas of Burkina Faso.

INTRODUCTION: Malaria still remains the most frequent parasitic disease on the world with, in 2022, 249 million cases and 608,000 deaths worldwide. Malaria control is compromised by the spread of the parasite's resistance to available antimalarials. The objective of our study is to characterize the Plasmodium falciparum resistance genes to common antimalarial drugs in semi-urban areas of Burkina Faso. MATERIALS AND METHODS: This is a prospective cross-sectional study whose collection took place from June to October 2021 and from June to October 2022 in five health facilities in Burkina Faso. The molecular analysis based on PCR-RFLP took place from January to June 2023 at Centre National de Recherche et de Formation (CNRFP) to determine resistance genes such as Pfcrt, Pfmdr1, Pfdhps, and Pfdhfr. RESULTS: A total of 150 samples were analyzed giving a prevalence of 46.67, 1.33, 0.67, 20, 82, and 4.67%, for Pfcrt 76 T, Pfmdr1 86Y, Pfdhps 437G, Pfdhfr 51I, Pfdhfr 59R, and Pfdhfr 108N mutations, respectively. There are no mutations observed Pfdhps 540E and Pfdhfr 164L positions. However, mutation on Pfdhfr 59R position was the most common. In addition, triple mutation (Pfdhps 437G + Pfdhfr 59R + Pfdhfr 108N) was found with a low frequency which is 0.67%. CONCLUSION: Surveillance of Plasmodium falciparum resistance markers to antimalarial drugs, remains one of the priorities in the context of the control or malaria elimination.

Plasmodium falciparum Genetic Diversity and Resistance Genotype Profile in Infected Placental Samples Collected After Delivery in Ouagadougou.

Purpose: Intermittent preventive treatment with sulfadoxine-pyrimethamine is widely used for the prevention of malaria in pregnant women in Africa. Known resistance cases of sulfadoxine-pyrimethamine during pregnancy need to be follow up to support IPTp implementation in Burkina Faso. However, data on the development and spread of resistance to this molecule are lacking. This study aimed to investigating the genetic diversity of P. falciparum and the mutation prevalence in the dhfr and dhps genes infected from postpartum infected placentas. Patients and Methods: This was a prospective and cross-sectional study conducted between April 2019 and March 2020 in four health districts of Ouagadougou capital city. From the placentas collected after delivery, P. falciparum detection and mps1 and msp2 polymorphism analysis were performed by nested PCR. The resistance profile was checked after analyzing the mutation point on dhfr and dhps genes. Results: PCR-positive samples were estimated at 96% for msp1 and 98% for msp2. The polymorphism analysis showed that the RO33 and 3D7 allelic families were the most widespread with 62.5% and 91.83%, respectively. Multiple infections by msp1 and msp2 were frequent with 12.50% and 92.92%, respectively. The prevalence of individual dhfr mutation point, 51I, 108A, and 59R, was 1.96, 15.68, and 7.84, respectively, and the dhps mutation point, 437G, was 3.92. There is no detected mutation at the point 164L and 540E. The triple (51I+108A+59R) in dhfr and quadruple (51I+108A+59R+ 437G) mutation were not found. Conclusion: The results showed that Plasmodium falciparum has a high genetic diversity of msp1 and msp2. This suggests that dhfr and dhps mutant genotypes are potential early warning factors in the increase in the sulfadoxine-pyrimethamine resistance.

[Epidemiology, microbiology, and outcomes of septicemia in children treated at the Charles de Gaulle University Pediatric Hospital in Burkina Faso]. / Aspects Épidémiologiques, Microbiologiques et Évolutifs des Bactériémies de L'enfant, au Centre Hospitalier Universitaire Pédiatrique Charles de Gaulle, Burkina, Faso. abdousal2000@yahoo.fr

The aim of this study is to describe the epidemiological and microbiological characteristics and outcome of children with septicemia at the Charles de Gaulle University Pediatric Hospital of Ouagadougou to help improve probabilistic antibiotic therapy in this type of infection. This retrospective descriptive study covered all the children from 0 to 15 years old seen over a period of 7 years in any hospital department with suspected bacteriemia and for whom the bacteriology laboratory performed a blood culture. During the study period, the laboratory received 842 requests for blood cultures and found 154 (18.3%) of them to be positive. Files for 81 of the 154 patients could be found and examined. The distribution according to age showed septicemia was most frequent among those aged 6-15 years (61.7% of the cases). Microbial identification showed the dominant species to be Salmonella enterica (serovars paratyphi and typhi) (58%) followed by Staphylococcus aureus (12.3%). The salmonella isolates had a high rate of resistance to amoxicillin, chloramphenicol and cotrimoxazole. Staphylococci were always sensitive to the antibiotics with which they were tested, although to a lesser extent for penicillin G. All patients routinely received antibiotic treatment, and 81.5% (n=66) were cured (5 children died and 10 left the hospital against medical advice). This study shows that the bacterial epidemiology of septicemia in our setting is dominated by salmonella. Trends in bacterial resistance to antibiotics showed that common antibiotics such as amoxicillin and cotrimoxazole are no longer acceptable as probabilist therapy here. They should be replaced in this type of infection by injectable third generation cephalosporin alone or combined with aminoglycosides.

Prevalence and Molecular Characterization of Extended Spectrum ß-Lactamase, Plasmid-Mediated Quinolone Resistance, and Carbapenemase-Producing Gram-Negative Bacilli in Burkina Faso.

The spreading of carbapenemase-producing gram-negative bacilli (GNB) must be considered as an "urgent" threat. The aim of this study was to determine the prevalence of extended spectrum ß-lactamase (ESBL), plasmid-mediated quinolone resistance (PMQR), and carbapenemase-producing GNB and to characterize the supporting genes in GNB specimens isolated from patients and healthy volunteers in Burkina Faso. From April to June 2016, carbapenemase-producing GNB screening was performed in 1,230 consecutive clinical specimens, and 158 fecal samples from inpatients and healthy volunteers without digestive pathology at Souro Sanou University Hospital, Bobo Dioulasso. Strains were identified by matrix-assisted laser desorption ionization-time of flight and antimicrobial susceptibility was tested with the disk diffusion method on Müller-Hinton agar. The presence of carbapenemase, ESBL, and PMQR genes was assessed by multiplex PCR. The molecular epidemiological study was performed using multilocus sequence typing analysis. From the 1,230 clinical samples, 443 GNB strains were isolated among which 4 (0.9%) were carbapenemase-producing isolates (Escherichia coli, n = 1; Acinetobacter baumannii, n = 3). Among the 158 fecal samples tested for carbapenemase-producing Enterobacteriaceae carriage, 13 (8.2%) were carbapenemase-producing isolates (E. coli, n = 4; Klebsiella pneumoniae, n = 6; A. baumannii, n = 2; Acinetobacter nosocomialis, n = 1; Acinetobacter bereziniae, n = 1). The strains from the two groups were resistant to broad-spectrum cephalosporins (100% for both), gentamicin (100% and 64.3%), levofloxacin (100% and 85.7%), and to amikacin (0% and 7.1%). The carbapenemase-encoding genes blaNDM-1, blaOxa-58, blaOxa-181, and blaVIM-2 were detected in clinical and in fecal samples. The majority (10/11) of the enterobacterial strains carried also blaCTX-M-15. The majority of the strains belonged to ST692 for E. coli, to ST147 for K. pneumoniae and to ST2 for A. baumannii. This study confirms the presence of carbapenemase-producing GNB in samples from patients and healthy volunteers. More effective active surveillance activities are needed.

[Criterium pencil mines as an alternative to platinum rods (Inoclic) in achievement of the antibiogram in agar medium for countries with limited resources?] / Les mines de crayon pour critérium comme alternative aux tiges de platine (Inoclic) dans la réalisation de l'antibiogramme en milieu gélosé pour les pays à ressources limitées ?

The realization of the antibiotic susceptibility test in agar is the routine bacteriological examination for the determination and monitoring of bacterial susceptibility to antibiotics. In this study, we report the comparative results between pencil leads for criterium, as an alternative to platinum rods in the realization of the antibiotic susceptibility test. METHODOLOGY: Experimental study evaluating the comparability of the results between Criterium and Inoclic mines (by counting bacterial cells on agar after 5 successive dilutions of reason 10 from a bacterial suspension obtained after piercing through a colony; by measuring the inhibition diameters of 4 ATCC reference bacterial strains on an antibiogram in an agar medium) and evaluating the sterility of the criterium mines by culturing them on enriched broth (heart - brain type). RESULTS: 42 bacterial strains were used for bacterial cell counting. The results were of the same order of magnitude (107 CFU/mL) between Inoclic and criterium mines, for all strains and at all dilutions. The antibiotic susceptibility tests performed for the 4 reference strains by the Inoclics and criterium mines all complied (100%) with the expected limits for determining their sensitivity profile to the antibiotics tested. Compared to the bacterial growth inhibition diameters on antibiotic susceptibility tests, no intra-operator variability was observed, while significant inter-operator variability (both with Inoclic and 0.5 mm criterium mines) was observed with some strains and for inhibition diameters greater than 10 mm. The enriched broth cultures (BCC) and their subculture carried out on 10 criterium mines from 5 different batches were negative. CONCLUSION: Criterium mines seem to be a serious and less expensive alternative to Inoclic for the realization of antibiotic susceptibility testing in our resource-limited countries.

Evaluation of pneumococcal meningitis clusters in Burkina Faso and implications for potential reactive vaccination.

BACKGROUND: To better understand how to prevent and respond to pneumococcal meningitis outbreaks in the meningitis belt, we retrospectively examined Burkina Faso's case-based meningitis surveillance data for pneumococcal meningitis clusters and assessed potential usefulness of response strategies. METHODS: Demographic and clinical information, and cerebrospinal fluid laboratory results for meningitis cases were collected through nationwide surveillance. Pneumococcal cases were confirmed by culture, polymerase chain reaction (PCR), or latex agglutination; strains were serotyped using PCR. We reviewed data from 2011 to 2017 to identify and describe clusters of ≥ 5 confirmed pneumococcal meningitis cases per week in a single district. We assessed whether identified clusters met the 2016 WHO provisional pneumococcal meningitis outbreak definition: a district with a weekly incidence of >5 suspected meningitis cases/100,000 persons, >60% of confirmed meningitis cases caused by Streptococcus pneumoniae, and >10 confirmed pneumococcal meningitis cases. RESULTS: Twenty pneumococcal meningitis clusters were identified, with a maximum weekly incidence of 7 cases and a maximum duration of 4 weeks. Most identified clusters (15/20; 75%) occurred before nationwide introduction of 13-valent pneumococcal conjugate vaccine (PCV13) in October 2013. Most cases were due to serotype 1 (74%), 10% were due to PCV13 serotypes besides serotype 1, and 8 clusters had >1 serotype. While 6 identified clusters had a weekly incidence of >5 suspected cases/100,000 and all 20 clusters had >60% of confirmed meningitis cases due to S. pneumoniae, no cluster had >10 confirmed pneumococcal meningitis cases in a single week. CONCLUSIONS: Following PCV13 introduction, pneumococcal meningitis clusters were rarely detected, and none met the WHO provisional pneumococcal outbreak definition. Due to the limited cluster size and duration, there were no clear instances where reactive vaccination could have been useful. More data are needed to inform potential response strategies.

Meningococcal carriage 7 years after introduction of a serogroup A meningococcal conjugate vaccine in Burkina Faso: results from four cross-sectional carriage surveys.

BACKGROUND: In the first 2 years after a nationwide mass vaccination campaign of 1-29-year-olds with a meningococcal serogroup A conjugate vaccine (MenAfriVac) in Burkina Faso, carriage and disease due to serogroup A Neisseria meningitidis were nearly eliminated. We aimed to assess the long-term effect of MenAfriVac vaccination on meningococcal carriage and herd immunity. METHODS: We did four cross-sectional studies of meningococcal carriage in people aged 9 months to 36 years in two districts of Burkina Faso between May 2, 2016, and Nov 6, 2017. Demographic information and oropharyngeal swabs were collected. Meningococcal isolates were characterised using whole-genome sequencing. FINDINGS: Of 14 295 eligible people, 13 758 consented and had specimens collected and laboratory results available, 1035 of whom were meningococcal carriers. Accounting for the complex survey design, prevalence of meningococcal carriage was 7·60% (95% CI 5·67-9·52), including 6·98% (4·86-9·11) non-groupable, 0·48% (0·01-0·95) serogroup W, 0·10% (0·01-0·18) serogroup C, 0·03% (0·00-0·80) serogroup E, and 0% serogroup A. Prevalence ranged from 5·44% (95% CI 4·18-6·69) to 9·14% (6·01-12·27) by district, from 4·67% (2·71-6·64) to 11·17% (6·75-15·59) by round, and from 3·39% (0·00-8·30) to 10·43% (8·08-12·79) by age group. By clonal complex, 822 (88%) of 934 non-groupable isolates were CC192, all 83 (100%) serogroup W isolates were CC11, and nine (69%) of 13 serogroup C isolates were CC10217. INTERPRETATION: Our results show the continued effect of MenAfriVac on serogroup A meningococcal carriage, for at least 7 years, among vaccinated and unvaccinated cohorts. Carriage prevalence of epidemic-prone serogroup C CC10217 and serogroup W CC11 was low. Continued monitoring of N meningitidis carriage will be crucial to further assess the effect of MenAfriVac and inform the vaccination strategy for future multivalent meningococcal vaccines. FUNDING: Bill & Melinda Gates Foundation and Gavi, the Vaccine Alliance.

Genotype Analysis of Clinical Candida albicans Isolates Using PCRs Targeting 25S rDNA and ALT Repeat Sequences of the RPS and Antifungal Susceptibility in Ouagadougou (Burkina Faso).

OBJECTIVE: Candida albicans is a yeast with multiple genotypes. It's a commensal fungus colonizing various sites. However, when the host's immune system weakens, it becomes pathogenic and is responsible for various lesions. In Burkina Faso, antifungal drugs are frequently used, particularly fluconazole, the most used systemic antifungal. This antifungal drug and other antifungal drugs are often used for self-medication or prescribed outside of antifungal susceptibility test results. These situations led to the emergence of Candida albicans strains resistant to antifungal drugs commonly used in Burkina Faso. The aim of this study was to determine the types of Candida albicans using PCRs targeting 25S rDNA and ALT repeat sequences of the RPS and to establish their azoles and polyenes susceptibility profile. MATERIAL AND METHODS: Antifungal susceptibility testing by disk diffusion method was performed in accordance with CLSI document M44-A for yeasts and the manufacturer's instructions. Candida albicans isolates were genotyped using specific PCR primers of the rDNA and RPS genes. RESULTS: Ten (10) RPS types of Candida albicans were found in our study: The most common RPS types are A3 (40.6%), A2 (24.0%) and A2/3 (14.6%) for genotype A, B2/3 (5.2%) for genotype B and C2 (3.2%) for genotype C. The Azole resistance, especially fluconazole (74.4%), was the most common with genotype A, including A3 (36.6%), A2 (18. 3%). Polyene resistance was rare with nystatin, only A3 (1.2%) resistant isolate to nystatin was observed. For amphotericin B, the highest observed resistance rates were A3 (11.0%) and A2/3 (8.5%) for the genotype A and B2 (10.0%), B3 (10.0%) and B2/3 (10.0%) for genotype B. CONCLUSION: Our study showed that Candida albicans resistance to azoles, especially to fluconazole, is an important phenomenon in Ouagadougou, and several genotypes RPS types are involved. Thus, fluconazole would not be an antifungal agent for first-line prescribing for treatment of candidiasis in Ouagadougou. This study will be continued to determine the molecular mechanisms involved in these antifungal resistances, for further research of new molecules with different action targets.

Triplex Real-Time PCR without DNA Extraction for the Monitoring of Meningococcal Disease.

Detection of Neisseria meningitidis has become less time- and resource-intensive with a monoplex direct real-time PCR (drt-PCR) to amplify genes from clinical specimens without DNA extraction. To further improve efficiency, we evaluated two triplex drt-PCR assays for the detection of meningococcal serogroups AWX and BCY. The sensitivity and specificity of the triplex assays were assessed using 228 cerebrospinal fluid (CSF) specimens from meningitis patients and compared to the monoplex for six serogroups. The lower limit of detection range for six serogroup-specific drt-PCR assays was 178⁻5264 CFU/mL by monoplex and 68⁻2221 CFU/mL by triplex. The triplex and monoplex showed 100% agreement for six serogroups and the triplex assays achieved similar sensitivity and specificity estimates as the monoplex drt-PCR assays. Our triplex method reduces the time and cost of processing CSF specimens by characterizing six serogroups with only two assays, which is particularly important for testing large numbers of specimens for N. meningitidis surveillance.

Early impact of 13-valent pneumococcal conjugate vaccine on pneumococcal meningitis-Burkina Faso, 2014-2015.

OBJECTIVES: We evaluate early impact of 13-valent pneumococcal conjugate vaccine (PCV13) on pneumococcal meningitis in Burkina Faso. METHODS: Nationwide surveillance gathered demographic/clinical information and cerebrospinal fluid (CSF) results for meningitis cases. Pneumococcal cases were confirmed by culture, polymerase chain reaction (PCR), or latex agglutination, and strains serotyped using PCR. We compared incidence (cases per 100,000) in the early post-PCV13 period (2014 and 2015) to average pre-PCV13 incidence (2011-2013). RESULTS: In 2015, age-specific pneumococcal meningitis incidences were 8.7 (<1 year), 2.4 (1-4 years), 6.5 (5-14 years), and 2.6 (≥15 years). Compared to 2011-2013, PCV13-serotype incidence among all ages decreased by 32% (95%CI: 23%-39%), with significant decreases among children aged <1 year (76%; 95%CI: 64%-84%) and 1-4 years (58%, 95%CI: 40%-71%). Among all ages, incidence of PCV13 serotypes besides serotype 1 decreased (68%; 95%CI: 59%-75%), but serotype 1 incidence did not. Incidence of non-PCV13 serotypes also decreased (47%; 95%CI: 29%-60%). Among children aged <1 year, serotypes 12F/12A/12B/44/46 (17%), 1 (12%), and 5 (10%) predominated. CONCLUSIONS: Following PCV13 introduction, PCV13-serotype meningitis incidence in young children significantly decreased. PCV13 impact on serotype 1 and disease in older children and adults requires continued monitoring.

Impact and effectiveness of pentavalent rotavirus vaccine in children <5 years of age in Burkina Faso.

BACKGROUND: Burkina Faso was one of the first African nations to introduce pentavalent rotavirus vaccine (RV5, RotaTeq) into its national immunization program in October 2013. We describe the impact and effectiveness of rotavirus vaccine on acute gastroenteritis (AGE) hospitalizations among Burkinabe children. METHODS: Sentinel hospital-based surveillance for AGE was conducted at four hospitals during December 2013 - February 2017. Demographic, clinical, and vaccination information was collected and stool specimens were tested by EIA. Trends in rotavirus AGE hospitalizations and changes in the proportion of AGE hospitalizations due to rotavirus were examined at two sentinel sites from January 2014 - December 2016. Unconditional logistic regression models using data from all 4 surveillance sites were used to calculate vaccine effectiveness (VE, defined as 1-odds ratio) by comparing the odds of vaccination among rotavirus AGE (cases) and non-rotavirus AGE (controls) patients, controlling for age, season, hospital site and socioeconomic factors. RESULTS: The proportion of AGE hospitalizations that tested positive for rotavirus declined significantly among children <5 years of age, from 36% (154/422) in 2014 to 22% (71/323, 40% reduction, p < .01) in 2015 and 20% (61/298, 44% reduction, p < .01) in 2016. Among infants, the percentage of AGE admissions due to rotavirus fell significantly from 38% (94/250) in 2014 to 21% (32/153, 44% reduction, p < .01) in 2015 and 17% (26/149, 54% reduction, p < .01) in 2016. The adjusted VE for full 3-dose series of RV5 against rotavirus hospitalization was 58% (95% [CI], 10%, 81%) in children 6-11 months of age and 19% (-78%, 63%) in children ≥12 months. CONCLUSION: Rotavirus hospitalizations declined after introduction of pentavalent rotavirus vaccine in children, particularly among infants. RV5 significantly protected against severe rotavirus gastroenteritis in infants, but effectiveness decreased in older children.