Evaluation of the Sysmex UF-4000i urine analyzer as a screening test to rule out urinary tract infection and reduce urine cultures. / Évaluation de l'analyseur Sysmex UF-4000i comme test de dépistage pour exclusion des infections urinaires et diminution des mises en culture

INTRODUCTION: Urinary tract infection (UTI) diagnosis by urine culture is time- and labor- consuming. In the Ibn Rochd microbiology laboratory, up to 70% of urine culture samples yield no growth or insignificant growth. OBJECTIVE: To evaluate the new generation of Sysmex UF-4000i fluorescence flow cytometry analyzer with a blue semiconducting laser as a method to rule out negative urine samples for UTI, in comparison of urine culture. MATERIAL AND METHODS: Flow cytometry and microbiological analysis were performed on 502 urine samples included in the study. We used ROC analysis to determine cutoff points at which optimal sensitivity and specificity are achieved for clinical use. RESULTS: Our results showed that bacteria count at a cut-off of 100/µL, and/or the leucocytes count ≥ 45/µL are the optimal indicator for positive culture results. At these cut off, bacteria sensitivity (SE), specificity (SP), Positive predictive value (PPV) and negative predictive value (NPV) were 97,3%, 95%, 87,8% and 98,8% respectively. For leucocytes, SE, SP, PPV and NPV were 99,1%, 95,8%, 88,6% and 99,7% respectively. DISCUSSION AND CONCLUSION: The bacterial and leucocytes counts generated by UF-4000i analysis may be useful in our context as a rapid screening to exclude UTI by reducing about 70% of urines cultures and then workload. Nevertheless, further validation is needed for different patient groups especially with urological disease or immunocompromised patients.

Serotype distribution and antimicrobial susceptibility of invasive Streptococcus pneumoniae isolates among adult and elderly population before and after introduction of pneumococcal conjugate vaccine in Casablanca, Morocco.

BACKGROUND: Streptococcus pneumoniae (S. pneumoniae), remains a major cause of mortality and morbidity worldwide. The objective of this study was to determine the trends of invasive pneumococcal diseases (IPD) in adult and elderly population in Casablanca (Morocco) before and after introduction of pneumococcal conjugate vaccine (PCV) by determining the distribution of pneumococcal serotypes and antibiotic resistance profile of isolated strains. METHOD: The proposed study is a retrospective laboratory-based surveillance of IPD in hospitalized adult (15-59 years old) and elderly (≥ 60 years old) patients in Ibn Rochd University Hospital Centre from 2007 to 2019 (13 years). All the 250 non-duplicate clinical invasive isolates from adult and elderly patients, confirmed as S. pneumoniae according to the laboratory standard identification procedures, are included in this study. RESULTS: A significant decrease of the overall incidence in IPD was observed only in adults from 0.71 to 0.54/100000 populations (P = 0.02) and to 0.47/100000 populations (P = 0.0137) in the early and mature post-vaccine period respectively compared to the pre-vaccine period. Our results also showed a significant reduction in the overall prevalence of vaccine serotypes from 28.17 to 6.90% (P = 0.0021) for the PCV-10 serotypes, and from 46.48 to 25.86% (P = 0.0164) for the PCV-13 serotypes only in the mature post-vaccine period (2015-2019). In parallel, the rate of non-vaccine serotypes did not significantly change in the early post-vaccine period (2011-2014) while it increased considerably from 54 to 74.14% (P = 0.0189) during the mature post-vaccine period. The rate of penicillin non-susceptible pneumococcal isolates decreased significantly from 23.94 to 8.77% (P = 0.02) in adult patients, and the rate of cotrimoxazole non-susceptible pneumococcal isolates significantly decreased from 29.58 to 8.77% in the early post-vaccine period (P = 0.003) and to 7.24% in the mature post-vaccine period (P = 0.0007). CONCLUSION: Although childhood vaccination has considerably reduced the incidence of IPD in adult population through the herd effect, IPD remain a real public health problem due to the alarming increase in non-vaccine serotypes (NVS) and the lack of herd effect among elderly population. The rate of antibiotic resistance was relatively low. Nevertheless, resistance constitutes a serious problem to the therapeutic arsenal due to the known capacity for genetic dissemination in the pneumococcus.

[Epidemiological profile of pertussis in infants in Casablanca from 2012 to 2019]. / Profil épidémiologique de la coqueluche du nourrisson à Casablanca de 2012 à 2019.

Pertussis is a real public health problem due to high neonatal morbidity rates and resurgence despite high vaccination coverage. The purpose of this study is to analyze the epidemiological profile of pertussis in infants hospitalized from 2012 to 2019. We conducted a retrospective, descriptive study over a 7-year and 8-month period from January 2012 to July 2019. It involved 500 infants admitted with clinical suspicion of pertussis. The average age of infants was 72 days, ranging from 28 days to 18 months; 75% of infants were less than 3 months old. The peak incidence was registered in 2012 and 2016, with a summer predominance (32%); 460 infants (92%) were not or incompletely vaccinated, 42.2% of whom were too young to be vaccinated. A probable contaminant in the entourage was found in 43,6% of cases. Whooping cough and cyanosis were the main reason for hospitalization (77.6%). Chest radiography objectified bronchial disease (25,4%) and alveolar foci (22.7%). Blood count performed in 410 infants showed hyperlymphocytosis in 67.5% of cases. Polymerase chain reaction (PCR) on nasopharyngeal sample collected from 206 infants was positive for Bordetella pertussis in 64% of cases; 118 PCR performed in mothers were positive in 47.7% of cases. All infants received Clarithromycin. Pertussis is a major cause of morbidity in infants in Casablanca. The prevention strategy is based on vaccination of family members of infants. However, vaccination of pregnant women appears to be more effective.

Evaluation of the Performances of the Rapid Test RESIST-5 O.O.K.N.V Used for the Detection of Carbapenemases-Producing Enterobacterales.

BACKGROUND: The emergence of carbapenemase-producing Enterobacterales (CPE) is a public health problem, requiring rapid and reliable diagnostic methods. The aim is to compare the new rapid immunochromatographic (IC) test: RESIST-5 O.O.K.N.V with PCR and the predictive model of EUCAST algorithm for the detection of CPE. METHODS: A longitudinal cross-sectional study was carried out in the bacteriology-virology laboratory of the Ibn Rochd-Casablanca University Hospital, from 1 February 2019 to 28 February 2020, concerning strains with reduced sensitivity to Ertapenem. The identification of bacterial species was carried out according to the standard criteria of microbiology and antibiogram according to CASFM-EUCAST 2019 recommendations. The sensitivity and specificity of the rapid IC test were calculated. RESULTS: The results of the new IC test showed a sensitivity and specificity of 100% for the detection of OXA-48 and NDM. These carbapenemases were detected simultaneously with a sensitivity and specificity of 100%. OXA-48 was the most common carbapenemas found (36%), followed by NDM (24%) and (13.4%) cases of OXA-48 and NDM coexistence. CONCLUSION: The rapid IC test could be a rapid and effective diagnostic tool for detecting the most common carbapenemases in our context, and to accelerate the implementation of adequate antibiotic therapy and infection control measures in patients with CPE infections.

Evaluation of the Sysmex UF-4000i urine analyzer as a screening test to rule out urinary tract infection and reduce urine cultures.

INTRODUCTION: Urinary tract infection (UTI) diagnosis by urine culture is time- and labor- consuming. In the Ibn Rochd microbiology laboratory, up to 70% of urine culture samples yield no growth or insignificant growth. OBJECTIVE: To evaluate the new generation of Sysmex UF-4000i fluorescence flow cytometry analyzer with a blue semiconducting laser as a method to rule out negative urine samples for UTI, in comparison of urine culture. MATERIAL AND METHODS: Flow cytometry and microbiological analysis were performed on 502 urine samples included in the study. We used ROC analysis to determine cutoff points at which optimal sensitivity and specificity are achieved for clinical use. RESULTS: Our results showed that bacteria count at a cut-off of 100/µL, and/or the leucocytes count ≥45/µL are the optimal indicator for positive culture results. At these cut off, bacteria sensitivity (SE), specificity (SP), Positive predictive value (PPV) and negative predictive value (NPV) were 97,3%, 95%, 87,8% and 98,8% respectively. For leucocytes, SE, SP, PPV and NPV were 99,1%, 95,8%, 88,6% and 99,7% respectively. DISCUSSION AND CONCLUSION: The bacterial and leucocytes counts generated by UF-4000i analysis may be useful in our context as a rapid screening to exclude UTI by reducing about 70% of urines cultures and then workload. Nevertheless, further validation is needed for different patient groups especially with urological disease or immunocompromised patients.

Epidemiological profile of Neisseria meningitidis in Casablanca, Morocco: 2010-2019.

Surveillance of invasive meningococcal diseases (IMD) must be carried out regularly and continuously in order to detect the emergence of strains of reduced susceptibility to antibiotics for therapeutic and prophylactic use and the appearance of new invasive clones. Molecular-typing approaches allow reliable traceability and powerful epidemiological analysis. This is an epidemiological study of Neisseria meningitidis causing meningitis in Casablanca, Morocco. The grouping was confirmed by PCR mainly on the isolates from cerebrospinal fluid (CSF). A total of 245 confirmed isolates of N .meningitidis were obtained between 2010 and 2019 of which 93 % are of group B. Overall, 24 % of all the isolates have a reduced susceptibility to penicillin G, but no resistance to penicillin G has been reported. All the isolated strains are susceptible to third-generation cephalosporins (3GCs). Genotyping by multilocus sequence typing (MLST) of a selection of 18 strains showed that the majority of isolates belong to the invasive clonal complex CC 32(9/18) followed by the CC 41/44(3/18).

Surgical wound infection caused by a multi drug resistant Streptococcus pneumoniae Serotype 19A after a total coloproctectomy with ileostomy.

Streptococcus pneumoniae (S. pneumoniae) colonizes asymptomatically the human nasopharynx. This pathogen is responsible for sinusitis, otitis media, pneumonia, bacteremia and meningitis. We report the case of a 35-year-old female patient who developed a surgical wound infection by a multi drug resistant S. pneumoniae serotype 19A after a total coloprotectomy. This first found in Morocco shows the implication of multidrug resistant S. pneumoniae in surgical wound infections.

Pertussis in infants, in their mothers and other contacts in Casablanca, Morocco.

BACKGROUND: In recent decades, there has been a marked increase in the number of reported cases of pertussis around the world, and pertussis continues to be a frequently occurring disease despite an effective childhood vaccination. This study aims to determine the role of household contacts of children diagnosed with pertussis in Casablanca Morocco. METHODS: From November 2015 to October 2017, children suspected of whooping cough that consulted Ibn Rochd University hospital at Casablanca with their household contacts were enrolled in the study. Nasopharyngeal (NP) samples of the suspected children were analyzed by culture and RT-PCR. For the household contacts, NP and blood samples were collected and analyzed by RT-PCR and specific detection of pertussis toxin antibodies by ELISA, respectively. RESULTS: During the study period, the survey was carried out on 128 infants hospitalized for pertussis suspicion and their families (N = 140). B. pertussis DNA was specifically detected in 73 (57%) samples, coexistence of B. pertussis and B. parapertussis DNA in 3 (2.3%) samples, coexistence of B. pertussis and B. holmesii DNA in 10 (7.81%) and only one (0.78%) sample was IS 481 RT-PCR positive without the possibility of determining the Bordetella species with the diagnostic tools used. Confirmations of Pertussis infection in household contacts by culture, RT- PCR and serology were 10, 46 and 39%, respectively. B. pertussis DNA was confirmed in the infants as well in their mothers in 38% of the cases. Co detection of B. pertussis and B. parapertussis DNA in 2% and co-detection of B. pertussis and B. holmesii DNA in 4%. B. holmesii DNA alone was detected in 5 NP samples of index cases and their mothers. CONCLUSIONS: The results of this study confirm that B. pertussis is still circulating in children and adults, and were likely a source of pertussis contamination in infants still not vaccinated. The use of RT-PCR specific for B. pertussis in the diagnosis of adults is less sensitive and should be associated with serologic tests to improve diagnosis of pertussis and contributes to preventing transmission of the disease in infants.

[Prevalence of coagulase-negative staphylococci in blood cultures at the Ibn-Rochd University Hospital in Casablanca]. / Prévalence des staphylocoques à coagulase négative dans les hémocultures au Centre Hospitalier Universitaire Ibn Rochd de Casablanca.

INTRODUCTION: Blood cultures are the best diagnostic tool for the detection of bacteremia. However, false positive results may lead to confusion about antibiotic regimens, putting the lives of patients at risk. The main purpose of this study was to assess the prevalence of coagulase negative Staphylococci (CoNS) as well as of Corynebacterium spp and Bacillus spp in the bags of blood culture analyzed in the microbiology laboratory at the Ibn-Rochd University Hospital in Casablanca. This prevalence was evaluated according to various Hospital Departments over the year 2016. METHODS: We conducted a descriptive, retrospective study by analysing the computerized database of the Laboratory of bacteriology and virology at the Ibn-Rochd University Hospital in Casablanca over a 12-month period from 1st January to 31st December 2016. Our study focused on bacteria forming part of the commensal flora (coagulase negative Staphylococcus, Corynebacteria spp and Bacillus spp). The blood culture bags were incubated in the automated blood culture system (Bactec FX). The identification of the germs from a positive culture was performed according to the standard techniques of bacteriology and susceptibility testing was performed according to EUCAST 2015. We conducted an analysis of the computerized database of KALISIL system (Netika) version (2.2.10.) of the Microbiology Laboratory at the Ibn-Rochd University Hospital in Casablanca. RESULTS: Out of 7959 requests for blood cultures obtained from 5801 patients addressed to the laboratory of bacteriology, 2491 were positive, of which 848, reflecting a rate of 34% of positive bags or 10.6% of the whole of bags received over the year 2016, were positive for coagulase negative Staphylococcus, 56 bags of blood cultures, reflecting a rate of 2.2%, were positive for Corrynébacteruim SP, followed by 60 bags of blood cultures, reflecting a rate of 2.4%, which were positive for Bacillus sp. The frequency of isolation of coagulase negative Staphylococcus compared to other bacteria according to Clinical Departments showed a higher frequency in the Paediatric Department (47.2%) followed by the Medicine Department (44.1%). CONCLUSION: This study shows that coagulase negative Staphylococci are the organisms most frequently isolated from blood cultures. They are a non-negligible cause of nosocomial infections, but they are also the most common blood culture contaminants.

[Meningitis caused by Streptococcus pneumoniae serotype 7a in an infant vaccinated with two doses of 13-valent pneumococcal conjugate vaccine: a case study]. / Méningite à Streptococcus pneumoniae sérotype 7A chez un nourrisson immunisé par deux doses du vaccin pneumococcique conjugué 13-valent: à propos d'un cas.

Pneumococcal meningitis is a global scourge. It is a major cause of morbidity and mortality. In Morocco, 13-valent pneumococcal conjugate vaccine (PCV13) was introduced into the National Immunization Program in October 2010 according to the immunization schedule 2 + 1 and replaced by PCV10 in July 2012, according to the same schedule. Despite the use of the PCV13, which is essential in the fight against pneumococcal disease, the emergence of new non-vaccine serotypes always results in meningitis in children, causing serious sequelae. We report the case of an infant vaccinated with two doses of PCV13 with meningitis caused by Streptococcus pneumoniae serotype 7a. The peculiarity of this case study lies in pneumococcal meningitis due to Streptococcus pneumoniae serotype 7a not included in the PCV13 in an infant immunized by 2 doses of PCV13. We here insist on the need and the importance of an observatory for pneumococcal meningitis and of a wide epidemiological study in order to determine the serotypes in Morocco after the introduction of PCV13 and then of PCV10.

Analysis of amino acid motif of penicillin-binding proteins 1a, 2b, and 2x in invasive Streptococcus pneumoniae nonsusceptible to penicillin isolated from pediatric patients in Casablanca, Morocco.

OBJECTIVES: This study aimed to investigate the nature of the amino acid motifs found in PBPs of Streptococcus pneumoniae isolates in invasive diseases from pediatric patients at Casablanca, Morocco. Five penicillin-susceptible (PSSP), ten penicillin-intermediate (PISP), and fifteen penicillin-resistant S. pneumoniae (PRSP) were studied by PCR-RFLP and DNA sequencing of the pbp1a, - 2b, and - 2x genes. RESULTS: There were no changes in the conserved motifs of PBP1a, PBP2b and PBP2x for PSSP strains. Substitution close to PBP1a conserved motifs were found in all PRSP isolates and six/five PISP. Analysis of PBP2b showed that all but one of the 10 PISP strains and all PRSP had substitutions. Substitution close to PBP2x motifs showed that all but three of the 10 PISP strains and all PRSP had substitutions in tow conserved motifs. A total of 6, 11 and 10 genotypes were found after analysis of pbp1a, pbp2b, and pbp2x, respectively. The penicillin-nonsusceptible S. pneumoniae isolated in Casablanca share most amino acid substitutions of those reported worldwide, but they occurred among pneumococci with low level resistance to b-lactams.

Molecular Characterization of Invasive Isolates of Neisseria meningitidis in Casablanca, Morocco.

Meningococcal epidemiology may change unpredictably, and typing of Neisseria meningitidis isolates is crucial for the surveillance of invasive meningococcal disease (IMD). Few data are available regarding the meningococcal epidemiology in countries of North Africa. We aimed to explore invasive meningococcal isolates from the Casablanca region in Morocco. We used whole-genome sequencing (WGS) to characterize 105 isolates from this region during the period of 2011 to 2016. Our data showed that the majority (n = 100) of the isolates belonged to serogroup B. Genotyping indicated that most of the isolates (n = 62) belonged to sequence type 33 of clonal complex 32. The isolates also showed the same PorA and FetA markers and clustered together on the basis of WGS phylogenetic analysis; they seemed to correspond to an expansion of local isolates in the Casablanca region, as reported for similar isolates in several other countries. These data suggest that serogroup B isolates may predominate in Morocco, which may have an important impact in the design of vaccination strategies.

[Healthcare-associated bacteraemia in intensive care units of Ibn Rochd University Hospital, Casablanca]. / Les bactériémies associées aux soins en réanimation au Centre hospitalier universitaire Ibn Rochd, Casablanca, Maroc.

Objective: This study was designed to determine the incidence of healthcare-associated bacteraemia in intensive care units (ICU) of Ibn Rochd University Hospital, Casablanca, Morocco, the frequency of the bacterial strains isolated and their antibiotic resistance patterns. Methods: A prospective longitudinal study was conducted between May and July 2015 on patients admitted to the Ibn Rochd University Hospital ICU. The Comité technique des infections nosocomiales et des infections liées aux soins (CITNILS) definition of bacteraemia was used. Micro-organisms were isolated and identified according to standard bacteriology techniques. Antibiotic susceptibility testing was performed according to CLSI 2013 guidelines. Results: A total of 720 patients were hospitalised during the study period; 48 (6.7%) acquired healthcare-associated bacteraemia with an incidence ranging from 7.3/1,000 HD to 13.7/1,000 HD. Fifty-four bacteria were isolated: A. baumannii and K. pneumoniae each accounted for 18.5% of isolates. 66.7% of A. baumannii strains were resistant to imipenem and 20% of K. pneumoniae strains were ESBL+ Conclusion: Healthcare-associated bacteraemia raises treatment problems due to multidrug-resistant bacteria, which is why regular surveillance is necessary to evaluate the time-course of these strains and the efficacy of prevention measures.

Epidemiology of pertussis in Casablanca (Morocco): contribution of conventional and molecular diagnosis tools.

BACKGROUND: Pertussis, a vaccine preventable disease, is still responsible of significant morbidity and mortality around the world, mostly in newborns. The aim of the present study was (1) to introduce pertussis surveillance in the major pediatric hospital of Casablanca (2) to analyze the prevalence of pertussis among children under 14 years of age and their entourage in Casablanca, Morocco. METHODS: This is a prospective and non-case controlled study, including children suspected of Pertussis admitted at the Abderrahim Harouchi Pediatric Hospital in Casablanca, from January 2013 to June 2015. Nasopharyngeal samples were obtained for Bordetella spp. culture and Real time PCR detection (RT-PCR) with specific primers of Bordetella spp., B. pertussis, B. parapertussis and B. holmesii. The detection of Bordetella spp. was also performed in some household contacts of the children suspected of pertussis. RESULTS: During the 2.5-years period, a total of 282 samples were collected from hospitalized children (156) and in some of their contacts (126). Among 156 samples from the children (from whom 57% were under 2 month of age), Bordetella DNA was detected in 61% (96/156) by RT-PCR. Among these positive samples, 91.7% (88/96) corresponded to B. pertussis DNA. Furthermore, in 39.5% (38/96) of the Bordetella positive samples, B. holmesii DNA was also detected. B. parapertussis DNA was detected in only one sample (1/156). Out of the 156 samples collected from the hospitalized children, only 48 were tested by culture, and 4 B. pertussis were isolated (8.3%). Among the 126 samples from the contacts of the children, mostly mothers (115 cases), Bordetella DNA was detected in 47% (59/126), 90% (53/59) being B. pertussis DNA. Moreover, B. holmesii DNA was also detected in 18.6% (11/59) of the Bordetella positive samples, and coexistence of B. pertussis and B. holmesii DNA in 36.5% (35/96). Two B. pertussis were isolated by culture performed on 43 samples of the contacts of the children (4.6%). CONCLUSIONS: This study highlights the circulation of B. pertussis but also of B. holmesii in Casablanca-Morocco with a high proportion of co-infections B. holmesii/B. pertussis in infants and their mothers, indicate that infection of non-vaccinated infants could be more associated with young parents. Moreover, the RT- PCR provides a sensitive and specific diagnosis of B. pertussis infections and distinguishes it from other Bordetella species, and is therefore suitable for implementation in the diagnostic laboratory.

Molecular characterization of penicillin non-susceptible Streptococcus pneumoniae isolated before and after pneumococcal conjugate vaccine implementation in Casablanca, Morocco.

BACKGROUND: Streptococcus pneumoniae is a major cause of morbidity and mortality worldwide, especially among children and the elderly. The ability to effectively treat pneumococcal infection has been compromised due to the acquisition of antibiotic resistance, particularly to ß-lactam drugs. This study aimed to describe the prevalence and molecular evolution of penicillin non-susceptible S. pneumoniae (PNSP) isolated from invasive diseases before and after pneumococcal conjugate vaccine implementation in Casablanca, Morocco. METHODS: Isolates were obtained from the Microbiology Laboratory of Ibn Rochd University Hospital Centre of Casablanca. Serogrouping was done by Pneumotest Kit and serotyping by the Quellung capsular swelling. Antibiotic susceptibility pattern was determined by disk diffusion and E-test methods. The PNSP were analyzed by pulsed-field gel electrophoresis (PFGE) and by genotyping of pbp1a, pbp2b, and pbp2x genes. RESULTS: A total of 361 S. pneumoniae isolates were collected from 2007 to 2014. Of these isolates, 58.7% were obtained before vaccination (2007-2010) and 41.3% after vaccination (2011-2014). Of the 361 isolates, 80 were PNSP (22.2%). Generally, the proportion of PNSP between pre- and post-vaccination periods were 31 and 13% (p = 0.009), respectively. The proportion of PNSP isolated from pediatric and adult (age > 14 years) patients decreased from 34.5 to 22.9% (p = 0.1) and from 17.7 to 10.2% (p = 0.1) before and after vaccine implementation, respectively. The leading serotypes of PNSP were 14 (33 vs. 57%) and 19A (18 vs. 14%) before and after vaccination among children. For adults, serotypes 19A (53%) and 23F (24%) were the dominant serotypes in the pre-vaccination period, while serotype 14 (22%) was the most prevalent after vaccination. There were 21 pbp genotypes in the pre-vaccination period vs. 12 for post-vaccination period. PFGE clustering showed six clusters of PNSP grouped into three clusters specific to pre-vaccination period (clusters I, II and III), two clusters specific to post-period (clusters V and VI) and a cluster (IV) that contained clones belonging to the two periods of vaccination. CONCLUSION: Our observations demonstrate a high degree of genetic diversity among PNSP. Genetic clustering among PNSP strains showed that they spread mainly by a restricted number of PNSP clones with vaccine serotypes. PFGE clustering combined with pbp genotyping revealed that vaccination can change the population structure of PNSP.

Prevalence of Acinetobacter baumannii bacteremia in intensive care units of Ibn Rochd University Hospital, Casablanca.

BACKGROUND AND OBJECTIVES: Acinetobacter baumannii bacteremia are grave because of the multi-resistance of the organism to antibiotics. This study aimed to determine the prevalence of A. baumannii isolated from blood cultures and to describe their antibiotic resistance patterns. MATERIALS AND METHODS: A retrospective longitudinal study was conducted on blood cultures between 2010 and 2014 from all Ibn Rochd University Hospital intensive care units; it was based on the exploitation of microbiology laboratory database (duplicates were excluded). Isolation and identification of A. baumannii were performed according to standard techniques of bacteriology and susceptibility testing as recommended by the CLSI. PCR was used to detect ß-Lactamase genes, blaOXA-51, blaOXA-23. RESULTS: Among the 4232 samples received at the laboratory, 2402 (56.8%) were positive. Negative coagulase Staphylococcus was isolated in 21.6% of cases followed by A. baumannii (9.2%), and K. pneumoniae (9.1%). A. baumannii strains were resistant to most antibiotics tested: imipenem (75.7%), ceftazidim (85.4%), cefotaxim (98.6%), gentamicin (78.1%), amikacin (63.5%) and ciprofloxacin (88.2%). All A. baumannii strains, resistant to carbapenem, tested were positive for blaOXA-51 genes and 87.5% expressed the blaOXA-23 genes. CONCLUSION: A. baumannii was the second germ frequently isolated from blood cultures in intensive care units. It was multi-resistant to antibiotics. The strengthening of hospital hygiene measures and surveillance of antibiotic resistance is needed to limit the spread of germs and to optimize the management of antibiotics.

Phenotypic and genotypic characterization of Streptococcus pneumoniae resistant to macrolide in Casablanca, Morocco.

In Morocco, the 13-valent pneumococcal conjugate vaccine (PCV-13) was introduced in the national immunization program (NIP) in October 2010 and replaced by the PCV-10 in July 2012. The present study aimed to determine the prevalence of erythromycin-resistant Streptococcus pneumoniae (ERSP) and to analyze the phenotypic and genotypic characteristics of these isolates in Casablanca, Morocco from January 2007 to December 2014. Isolates were obtained from the Microbiology Laboratory of Ibn Rochd University Hospital Centre of Casablanca. Serogrouping was done using Pneumotest Kit and serotyping by the Quellung capsular swelling. Antibiotic susceptibility pattern was determined by disk diffusion and Etest methods. A total of 655S. pneumoniae isolates were collected from 2007 to 2014 from pediatric and adult patients. Fifty-five percent of these isolates were from invasive pneumococcal diseases. Of the 655 isolates, 92 (14%) were ERSP. Globally, the proportion of ERSP from 2007 to 2010 (before vaccination) and from 2011 to 2014 (after vaccination) were 11.6% and 17.2% (p=0.04), respectively. Of the 92 ERSP, 89%, 4% and 7% displayed constitutive MLSB (resistance to macrolide, lincosamide and streptogramin B), inducible MLSB, and M phenotype (resistance to macrolide only), respectively. ERSP genotypic analysis showed that 90.2% carried the ermB gene, 6.5% the mefE gene, and 3.3% both the genes (ermB+mefE). The most prevalent ERSP serotypes were 6B, 19F and 23F before vaccination and 19F, 6B, 6A and 23F after vaccination. Erythromycin resistance among S. pneumoniae is relatively high in Casablanca. The contribution of PCVs to the reduction in antibiotic use is encouraging but this should be accompanied by a rational use of antibiotic.

A duplex real-time PCR for the detection of Streptococcus pneumoniae and Neisseria meningitidis in cerebrospinal fluid.

INTRODUCTION: Acute bacterial meningitis is one of the most severe infectious diseases. Rapid, accurate, and inexpensive diagnosis of bacterial meningitis is crucial for patient management. This study describes a duplex real-time (RT) PCR assay for detection of Neisseria meningitidis and Streptococcus pneumoniae in the cerebrospinal fluid (CSF) for meningitis diagnosis using SYBR Green-based RT-PCR method coupled with melting curve analysis. METHODOLOGY: We used SYBR Green-based RT-PCR method coupled with melting curve analysis to detect S. pneumoniae and N. meningitidis in CSF samples. The sensitivity, specificity, and limit of detection were determined. The gold standard for routine tests of CSF analysis is direct examination, culture, and/or latex agglutination. The assay was evaluated on 132 CSF samples to measure clinical sensitivity. RESULTS: A duplex RT-PCR assay for N. meningitidis and S. pneumoniae detection in CSF was evaluated. Two peaks at different melting temperatures (87.5°C and 85.5°C) for N. meningitidis and S. pneumoniae, respectively, were obtained. The sensitivity of RT-PCR was 100% (95% confidence limits [CI] = 82.4-100) for N. meningitidis and 100% (95% CI = 85.1-100) for S. pneumoniae. Specificity was the same (100%) for the bacteria (95% CI = 88.6-100). The percentage of cases accurately diagnosed with meningitis caused by N. meningitidis and S. pneumoniae increased to 50.7% and 28.6%, respectively, when RT-PCR was added to the standard microbiologic methods. CONCLUSIONS: Duplex RT-PCR and melting curve analysis with SYBR Green is an inexpensive, sensitive, and specific method to rapidly diagnose bacterial meningitis. Accurate identification of the bacterial causative agents will improve patient management and epidemiological investigations.

Invasive pneumococcal disease among children younger than 5 years of age before and after introduction of pneumococcal conjugate vaccine in Casablanca, Morocco.

OBJECTIVES: The purpose of this study was to compare the incidence rate of invasive pneumococcal disease, the rates of antibiotic resistance and serotype distribution among children ≤5 years old before and after PCVs introduction in Casablanca, Morocco. METHODS: This study was conducted at the Ibn Rochd University Hospital Centre of Casablanca during two periods encompassing pre-and post-implementation of PCVs, respectively from January 2007 to October 2010 and from January 2011 to December 2014. All the non-duplicate invasive S. pneumoniae isolates recovered during the study periods were included. RESULTS: There were 136 cases of IPD, 91 before and 45 after PCVs introduction. The greatest decrease in incidence rate of IPD occurred in children ≤ 2 years of age declining from 34.6 to 13.5 per 100,000 populations (p<0.0001) before and after vaccination, respectively. The incidence rate of PCV-7, PCV-10 non-PCV-7 and PCV-13 non-PCV-10 serotypes decrease significantly from 18.0 to 4.6, from 5.7 to 1.3 and from 5.7 to 0.8/100,000 population (p<0.001) in the same age, respectively. CONCLUSION: Shifts in the distribution of IPD serotypes and reductions in the incidence rate of disease suggest an effective reduction of the burden of IPD in children, but continued high quality surveillance is critical to assess the changes in serotype distributions.

[Isolation of Bordetella trematum from bacteremia]. / Isolement de Bordetella trematum au décours d'une bactériémie.

The species Bordetella trematum was first described in 1996. Currently only eleven cases were published. We describe the first case of Bordetella trematum issued from bacteremia with a patient who has severe burns in Morocco. The identification was not possible by conventional microbiological methods where the resort to 16S ARNr sequencing. The use of molecular methods, including sequencing of the 16S ARNr, is currently an essential complementary tool to identify microbiological pathogens.