Evaluation of GeneXpert MTB/RIF system performances in the diagnosis of extrapulmonary tuberculosis.

BACKGROUND: Tuberculosis represents a serious public health problem and a significant diagnostic and therapeutic challenge worldwide. Molecular diagnostic techniques are crucial in the World Health Organization's new tuberculosis control strategy. This study aims to evaluate the performance of GeneXpert MTB/RIF (Cepheid Sunnyvale, CA, United States) in diagnosis of extra-pulmonary tuberculosis then compare it's performance in detecting Rifampicin resistance to GenoType MTBDRplus (HAIN Life Sciences, Nehren, Germany). METHODS: Samples from pulmonary and/or extra-pulmonary origins were analysed in a 21 months retrospective study. Samples were sent to the bacteriology laboratory for Mycobacterium tuberculosis detection using conventional bacteriological and molecular methods (GeneXpert MTB/RIF and MTBDRplus). Sensitivity and specificity were calculated for the stained smear and GeneXpert according to culture (Gold Standard) as well as for GeneXpert MTB/RIF in both negative and positive microscopy tuberculosis cases. Data's statistical analysis was performed with SPSS13.0 software. RESULTS: Seven hundred fourteen patients' samples were analysed; the average age was 47.21 ± 19.98 years with a male predominance (66.4%). Out of 714 samples: 285 were from pulmonary and 429 were from extra-pulmonary origins. The positivity rates for microscopy, GeneXpert MTB/RIF and culture were 12.88, 20.59 and 15.82%, respectively. These rates were 18.9, 23.85 and 20.35% for pulmonary samples and 9.71, 18.41 and 12.82% for extra-pulmonary samples, respectively. The sensitivity and specificity of GeneXpert MTB/RIF were almost the same in both pulmonary and extra-pulmonary samples (78.2 and 90.4%) and (79,3 and 90.3%) respectively. Rifampicin resistance rate found by GeneXpert MTB/RIF was 0.84%. Comparison of Rifampicin resistance obtained by GeneXpert MTB/RIF and Genotype MTBDRplus, showed 100% agreement between the two techniques for studied samples. CONCLUSIONS: This confirms GeneXpert MTB/RIF advantage for tuberculosis diagnosis, particularly extra-pulmonary tuberculosis with negatively stained smear. The performance of GeneXpert and Genotype MTBDRplus are similar in detection of Rifampicin resistance. However, variability of detection performance according to tuberculosis endemicity deserves more attention in the choice of screening techniques of Rifampicin resistance, hence the interest of conducting comparative studies of detection performance under low and medium endemicity on large samples of tuberculosis populations.

Peritoneal tuberculosis, an underestimated diagnosis: a case report.

Tuberculosis is an infectious disease that most often affects the lungs, caused by human-to-human transmission of Mycobacterium tuberculosis. Peritoneal tuberculosis is an extra-pulmonary form of the disease that usually manifests as an ascitic syndrome, with or without fever, in a context of altered general condition, often in endemic areas. The diagnosis of peritoneal tuberculosis is not always easy, as the clinical signs are often insidious and unspecific. We report a case of peritoneal tuberculosis in an 18-year-old female, who had presented for 10 days with a progressive increase in abdominal volume associated with vomiting and diarrhoea.

Bacteriological aspects of bacteremia in the intensive care unit of the Mohammed V Military Hospital: 10 months prospective study.

Introduction: Bacteremia is responsible for high rates of morbidity and mortality. The increasing prevalence of multidrug-resistant (MDR) bacteria in intensive care units (ICU) is a growing concern. Hence, prior knowledge of bacterial epidemiology and resistance phenotypes is required to optimize these infections' management. The objective of this study was to determine the epidemiological profile of bacteremia in ICU settings, as well as the place occupied by MDR bacteria in these infections. Methods: It is a prospective study carried out over 10 months on episodes of bacteremia in the ICU of Mohammed V Military Teaching Hospital (Rabat, Morocco). Microorganism growth was detected using fluorescent technology, species identification was based on morphological and biochemical characteristics. Antimicrobial susceptibility testing was performed following the recommendations of the Antibiogram Committee of the French Society of Microbiology (CA-SFM) and the European Committee on Antimicrobial Susceptibility Testing (EUCAST). Results: Among 504 hospitalized patients, sixty-one (12.1%) presented at least one episode of bacteremia. Forty patients (65.6% of bacteremic patients) presented at least one episode of bacteremia due to MDR bacteria. Male gender, cardiovascular diseases, diabetes and previous hospitalization were significant risk factors for the acquisition of MDR bacteremia. Isolated bacteria were mainly Gram-negative bacilli (GNB) (n = 62; 68.9%) dominated by Acinetobacter baumannii (n = 19; 21.1%) and Klebsiella pneumoniae (n = 16; 17.8%). MDR bacteria were represented by multi-resistant Acinetobacter baumannii (n = 19; 44.2%), extended-spectrum beta-lactamases-producing Enterobacterales (n = 9; 20.9%) and carbapenem-resistant Enterobacterales (n = 7; 16.3%). Carbapenems (n = 40; 65.6%), Aminoglycosides (n = 32; 52.5%) and Polypeptides (n = 24; 39.3%) were the most used antimicrobials. Mortality rates were 66.6% (n = 40) and 85% (n = 43) in patients with non MDR bacteremia and MDR bacteremia respectively. Conclusion: Limiting the spread of MDR bacteria and improving the management of bacteremic patients require continuous monitoring of bacteremia as well as adapting the therapeutic and preventive strategy.

Multidrug-resistant community-acquired urinary tract infections in a northern region of Morocco: epidemiology and risk factors.

INTRODUCTION: Urinary tract infection is very common and is the second most frequent reason for consultation in office-based practice. The incidence varies from one country to another. The diffusion of MDR in the community complicates therapeutic management. The objective of this study was to describe the bacterial epidemiology and to determine the risk factors for the acquisition of MDR in community urinary tract infections in our region. METHODS: This was a retrospective case-control study conducted in the bacteriology laboratory of the Mohammed V Military Teaching Hospital over an 8-month period from 01 October 2015 to 31 May 2016. Cases were defined as patients with community-acquired urinary tract infection with MDR and controls were defined as patients with a urinary tract infection without MDR. RESULTS: Out of 373 isolates, enterobacteria represented 80%. E. coli represented 59.2%, followed by K. pneumoniae at 15%. The rate of MDR represented 13.4% of which ESBL enterobacteria represented 12.1%. Univariate analysis showed a statistically significant association between male sex (p=0.001), age >65 years (p=0.007), urban origin (p=0.003), previous hospitalization within 3 months (p=0.001) and antibiotic therapy within 6 months (p=0.001) with MDR community-acquired urinary tract infection. On the other hand, multivariate analysis by logistic regression showed that age >65 years (OR=8.4, CI: 2.1-42), previous hospitalization within 3 months (OR=13.4, CI: 3.3-140.2) and antibiotic therapy within 6 months (OR=9.2, CI: 4.1-60.1) were significantly associated to MDR community-acquired urinary tract infection. CONCLUSIONS: The increase in resistance to enterobacteria in the community prompts a review of the list of antibiotics prescribed for probabilistic management of these infections in our region.

[Prevalence of male infertility in a university hospital in Morocco]. / Prévalence de l´infertilité masculine dans un hôpital universitaire au Maroc.

INTRODUCTION: infertility in couples has become a public health problem in recent years. It can be related to a problem in the male, female or both. Men infertility accounts for 40% of cases. In Morocco, most studies have focused on the causes and risk factors for male infertility. The purpose of our study was to assess the prevalence of male infertility and semen parameters in infertile men or in men at high risk of developing infertility in a tertiary hospital in Rabat. METHODS: we conducted an analysis of 482 patients referred for evaluation of infertility in the couple or as part of preoperative assessment of varicocele or testicular ectopia. Demographic data, risk factors for infertility, primary or secondary infertility were recorded for each patient. Semen parameters were assessed and interpreted according to WHO standards updated in 2010 based on studies of factors associated with their disturbance. RESULTS: the average age of patients was 35.35±8.81 years. Primary infertility was found in 61.8% of cases. The most common risk factors for infertility were tobacco followed by varicocele and infection. Spermogram was altered in 53.1% of cases. The most common abnormality was sperm vitality issues (36.9%) followed by spermatic concentration (29.7%) and morphology (29.3%). Age was the unique factor which had a significant impact on spermogram (p=0.002). Abnormalities in mobility were detected in patients ≥31 years, sperm vitality issues in patients aged 34 years, abnormalities in morphology in patients aged 35 years and concentration in patients aged 37 years. Azoospermia was found in 16.4% of cases and was mainly associated with primary infertility. Oligo-astheno-teratozoospermia was the most common association (26.2%). CONCLUSION: male infertility is common in our context. The major risk factor is age. Mobility is the earliest parameter achieved.

Update on the resistance of Escherichia coli isolated from urine specimens in a Moroccan hospital: a review of a 7-year period.

INTRODUCTION: Escherichia coli is the most commonly isolated species in both community and healthcare-associated infections. Our study's purpose was to determine the rates of antibiotic resistance of E. coli isolates in hospital and community populations, track the kinetics of resistance rates of E. coli isolates to major antibiotics, particularly those prescribed for urinary tract infections and study the occurrence and evolution of multi-resistant phenotypes. METHODS: We conducted a retrospective study at the Bacteriological Department of the Mohammed V Military Hospital of Instruction, over a period of 7 years. All isolates of E. coli from inpatients and outpatients were included. Identification of bacterial isolates was based on culture, morphological and biochemical identification characteristics. Antibiotic susceptibility was studied using the Mueller Hilton agar diffusion method by using OXOID® type antibiotic discs and interpreted according to the recommendations of EUCAST/CA-SFM 2019. RESULTS: The rate of resistance of E. coli isolates to 3rd generation cephalosporins, imipenem and fluoroquinolones was 12%, 1% and 34%, respectively. The difference between the resistance rates of inpatient and outpatient E. coli isolates was statistically significant for most antibiotics (p<0.05). The rate of extended-spectrum beta-lactamase phenotype (ESBL) was 6.73%. The carbapenemase phenotype was 1.25%. The ESBL phenotype rate increased from 3% in 2012 to 11.16% in 2018. CONCLUSIONS: The progression of the ESBL phenotype in both hospital and community settings, due to the increase in the resistance rate to 3rd generation cephalosporin, is prompting a review of the strategy for the therapeutic management of urinary tract infections with these molecules as probabilistic treatment.

Intensive care unit-acquired Acinetobacter baumannii infections in a Moroccan teaching hospital: epidemiology, risk factors and outcome.

INTRODUCTION: The objective of this study was to examine the epidemiology, risk factors and outcome associated with Acinetobacter baumannii infections in the intensive care units (ICUs) in a Moroccan teaching hospital. METHODS: This is a matched case-control study conducted as a joint collaboration between the clinical Bacteriology department and the two ICUs of Mohammed V Military Teaching Hospital from January 2015 to July 2016. RESULTS: Among 964 patients hospitalized in the ICUs, 81 (8.4%) developed A. baumannii infections. Multivariate logistic regression analysis identified the following independent risk factors for ICU-acquired A. baumannii infections: ICU stay ≥14 days (odds ratio (OR)=6.4), prior use of central venous catheters (OR=18), prior use of mechanical ventilation (OR=9.5), duration of invasive procedures ≥7 days (OR=7.8), previous exposure to imipenem (OR=9.1), previous exposure to amikacin (OR=5.2), previous exposure to antibiotic polytherapy (OR=11.8) and previous exposure to corticotherapy (OR=5). On the other hand, the admission for post-operative care was identified as a protective factor. The crude mortality in patients with A. baumannii infection was 74.1%. Multivariate analysis showed that septic shock (OR=19.2) and older age (≥65 years) (OR=4.9) were significantly associated to mortality risk in patients with A. baumannii infection. CONCLUSION: Our results show that shortening the ICU stay, rational use of medical devices and optimizing antimicrobial therapy could reduce the incidence of these infections. Elderly patients and those with septic shock have a poor prognosis. These findings highlight the need for focusing on the high-risk patients to prevent these infections and improve clinical outcome.

Clonal diversity and detection of carbapenem resistance encoding genes among multidrug-resistant Acinetobacter baumannii isolates recovered from patients and environment in two intensive care units in a Moroccan hospital.

BACKGROUND: Carbapenem-resistant Acinetobacter baumannii has recently been defined by the World Health Organization as a critical pathogen. The aim of this study was to compare clonal diversity and carbapenemase-encoding genes of A. baumannii isolates collected from colonized or infected patients and hospital environment in two intensive care units (ICUs) in Morocco. METHODS: The patient and environmental sampling was carried out in the medical and surgical ICUs of Mohammed V Military teaching hospital from March to August 2015. All A. baumannii isolates recovered from clinical and environmental samples, were identified using routine microbiological techniques and Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry. Antimicrobial susceptibility testing was performed using disc diffusion method. The carbapenemase-encoding genes were screened for by PCR. Clonal relatedness was analyzed by digestion of the DNA with low frequency restriction enzymes and pulsed field gel electrophoresis (PFGE) and the multi locus sequence typing (MLST) was performed on two selected isolates from two major pulsotypes. RESULTS: A total of 83 multidrug-resistant A. baumannii isolates were collected: 47 clinical isolates and 36 environmental isolates. All isolates were positive for the blaOXA51-like and blaOXA23-like genes. The coexistence of blaNDM-1/blaOXA-23-like and blaOXA 24-like/blaOXA-23-like were detected in 27 (32.5%) and 2 (2.4%) of A. baumannii isolates, respectively. The environmental samples and the fecally-colonized patients were significantly identified (p < 0.05) as the most common sites of isolation of NDM-1-harboring isolates. PFGE grouped all isolates into 9 distinct clusters with two major groups (0007 and 0008) containing up to 59% of the isolates. The pulsotype 0008 corresponds to sequence type (ST) 195 while pulsotype 0007 corresponds to ST 1089.The genetic similarity between the clinical and environmental isolates was observed in 80/83 = 96.4% of all isolates, belonging to 7 pulsotypes. CONCLUSION: This study shows that the clonal spread of environmental A. baumannii isolates is related to that of clinical isolates recovered from colonized or infected patients, being both associated with a high prevalence of the blaOXA23-like and blaNDM-1 genes. These findings emphasize the need for prioritizing the bio-cleaning of the hospital environment to control and prevent the dissemination of A. baumannii clonal lineages.

First Whole-Genome Sequences of Two Multidrug-Resistant Acinetobacter baumannii Strains Isolated from a Moroccan Hospital Floor.

This report describes the whole-genome shotgun sequences of two multidrug-resistant Acinetobacter baumannii strains, ABE8_07 and ABE12_M, isolated from a Moroccan hospital floor. These two genome sequences will initiate the study and characterization of the Acinetobacter baumannii genome in Morocco.