Evaluation of a Molecular Test for Detection of Mycobacterium tuberculosis Isolates Resistant to Rifampicin and Isoniazid.

BACKGROUND: Multidrug-resistant tuberculosis (MDR-TB) is increasing worldwide and is a major cause of death in many countries. It has become a major challenge for national tuberculosis control programs. Therefore, rapid identification of MDR strains of Mycobacterium tuberculosis and monitoring of their transmission could contribute significantly to the fight against tuberculosis. The GenoType MTBDRplus assay has been recommended by the World Health Organization to identify rifampicin (RIF)- and isoniazid (INH)-resistant M. Tuberculosis isolates. The objectives of this study were to evaluate the performance of the GenoType MTBDRplus test in the detection of rifampicin and isoniazid resistance of M. tuberculosis isolates in a Moroccan hospital and then to determine the frequency of mutations associated with resistance to these two major anti-tuberculosis drugs. METHODS: This is a retrospective study conducted at the bacteriology department of the Mohammed V military hospital over a period of one year from 01/01/2018 to 12/31/2019. A total of 92 isolates of M. tuberculosis from pulmonary and extra-pulmonary specimens were evaluated for drug susceptibility by MGIT™ 960 AST system and compared to the GenoType MTBDRplus assay. The MGIT™ 960 AST system was used as the gold standard for the evaluation of the GenoType MTBDRplus assay. RESULTS: Sensitivity and specificity of the GenoType MTBDRplus assay for the detection of RIF-resistant M. tuberculosis isolates were 83.33% and 100%, respectively. Its sensitivity and specificity for the detection of INH-resistant M. tuberculosis were 88.23% and 100% respectively. The concordances of the GenoType MTBDRplus assay and the MGIT™ 960 AST system for the detection of sensitivity to RIF and INH were 99% (1/92) and 98% (2/92), respectively. Among the five RIF-resistant isolates, the MUT3 mutation in the rpoB gene (codon S531L mutation) was present in 80% of isolates, whereas mutations in the rpoB MUT1 gene were present in only one (20%) RIF-resistant isolate. INH resistance was detected in 15 isolates, of which nine isolates (60%) had specific mutations of the katG gene (codon S315T1) and conferred a high level of resistance to INH. CONCLUSIONS: The results of this study have shown that the GenoType MTBDRplus test has a high sensitivity and specificity for the detection of resistance to RIF and INH.

A Case of Colonic Tuberculosis Revealed by Polymerase Chain Reaction.

BACKGROUND: The authors report a case of tumor-like colonic tuberculosis revealed by PCR in a 32-year-old patient with a low-level peritoneal effusion on CT scan with negative histological study on colonic biopsy. METHODS: The colonic biopsy received at the laboratory after grinding in a porcelain mortar, was the object of a molecular study by GeneXpert MT/RIF (Cepheid, Sunnyvale, CA, USA) using the automated real-time PCR technique and a conventional study based on Ziehl-Nielsen staining and culture on Lowenstein-Jensen® solid medium (LJ) and Mycobacteria Growth Indicator Tube (MGIT®) liquid medium. RESULTS: The patient was a 32-year-old male without any personal or family history of tuberculosis and without signs of tuberculosis impregnation. He had a story of ingestion of non-pasteurized dairy products including milk and cheese. For 45 days he had constipation with abdominal pain and feeling of heaviness. Physical examination of the patient revealed abdominal tenderness without adenopathy. The laboratory workup showed a normal blood count, CRP, liver and kidney function tests. The HIV test was negative. Medical imaging revealed a low-level peritoneal effusion that could not be punctured. Colonoscopy showed a thickening of the colon. The colonic biopsy, after crushing and sonication, was searched for the Mycobacterium tuberculosis complex by both molecular biology and conventional methods. Molecular research by GeneXpert MTB/RIF (Cepheid, Sunnyvale, CA, USA) using the automated real-time PCR technique, revealed the presence of the Mycobacterium tuberculosis complex without detection of rifampicin resistance. On the other hand, the direct examination after special Ziehl-Nielsen staining was positive (Figure 2) and the cultures on Lowenstein-Jensen® solid medium (LJ) and Mycobacteria Growth Indicator Tube (MGIT®) liquid medium were also positive after two and three weeks, confirming the molecular diagnosis. The histology study showed moderate non-specific chronic colitis with no histological arguments for tuberculosis or malignancy. The patient was placed on curative tuberculosis treatment according to the national protocol, with a favorable clinical-radiological course. CONCLUSIONS: Colonic tuberculosis is a disease that may mimic many other diseases; therefore, a correct approach is necessary for the correct diagnosis and treatment.

Cytological examination of cerebrospinal fluid: Sysmex UF-1000i versus optical microscopy.

We evaluated the body fluid module on Sysmex UF-1000i (UF-1000i-BF) for analysis of white blood cell (WBC) and red blood cell (RBC) in cerebrospinal fluid. We collected 93 cerebrospinal fluid samples and compared the results of the UF-1000i-BF mode with the Fast-Read 102 disposable counting cell. Results shows a good correlation between the UF-1000i and the microscopic examination. The concordance percentage is 99.06% for white blood cells and 85.18% for red blood cells. The UF-1000i-BF mode offers rapid and reliable total WBC and RBC counts for initial screening of cerebrospinal fluid, and can improve the workflow in a routine laboratory.

[Prevalence and role of IgG anti-Chlamydia trachomatis in a population of infertile men in Morocco]. / Prévalence et rôle des IgG anti-Chlamydia trachomatis chez une population d'hommes infertiles au Maroc.

INTRODUCTION: Infertility is a global public health problem that affects 15% of couples of childbearing age. Male infertility is involved in 20 to 50% of cases. These figures are sharply increasing around the world. Several factors may be responsible for this infertility with especially hormonal, genetic, toxic or infectious factors. The latter are dominated mainly by Chlamydia infection. Among the most serious complications of this infection are infertility related to urethritis, epididymitis and irreversible total azoospermia in men and tubal obstructions and ectopic pregnancies in women. STUDY OBJECTIVE: To determine the prevalence of IgG anti-Chlamydia trachomatis in men consulting for infertility and the association between previous contact with this bacterium and the impairment of sperm quality and sperm function. MATERIAL AND METHODS: Prospective study over 26months of 143 patients referred to the service for infertility assessment of the couple. Demographic data, primary or secondary character of infertility, risk factors (tobacco, inguinal hernia, varicocele and history of urogenital infections), semen parameters (volume, mobility, pH, vitality and morphological abnormalities) were studied as well as the determination of the anti-C. trachomatis IgG titer. The prevalence of Chlamydia infection and the association of the infection and alteration of the various parameters of the semen were analyzed. RESULTS: The average age of patients was 38.5±8.55. Infertility was primary in 72% of patients. Among the patients, 54.5% had an abnormal spermogram. Chlamydia IgG antibodies were positive in 37.1% of patients whose 58.5% had abnormal spermogram. Analysis of sperm parameters of patients with and without IgG C. trachomatis showed an altered vitality in Chlamydia positive patients with an OR at 2.41, P=0.02, (95% CI: 1.15-5.06). CONCLUSION: The prevalence of Chlamydia infection is high in infertile male. C. trachomatis IgG antibodies may be associated with an alteration of spermatozoa vitality without significant impairment of other semen parameters. LEVEL OF EVIDENCE: 3.

[Visceral leishmaniasis, pemphigus and immunosuppressive treatment: case report from Morocco]. / Leishmaniose viscérale, pemphigus et traitement immunosuppresseur: une observation au Maroc.

Atypical forms of visceral leishmaniasis associated with immunosuppressive treatment are difficult to diagnose and cause high mortality. The purpose of this report is to describe a case involving a 42-year-old patient living in a leishmaniasis-endemic area, who was undergoing immunosuppressive treatment using corticosteroids and methotrexate for pemphigus. Despite clinical and laboratory findings consistent with visceral leishmaniasis, detection of Leishmania bodies was a coincidental finding of cytological examination of bone marrow during workup for pancytopenia and associated clinical signs. This case argues in favor of systematic screening for this opportunistic parasitic disease before undertaking immunosuppressive treatment in patients presenting risk factors and consistent clinical/laboratory findings.

Shewanella putrefaciens: A cause of bacteremia not to neglect.

Shewanella putrefaciens is a Gram-negative bacillus and marine pathogen that rarely causes disease in humans. We report the first Moroccan case of multidrug-resistant Shewanella putrefaciens bacteremia and describe the bacteriological, clinical, and antibiogram characteristics of this isolate, which was repeatedly isolated from the blood of a 66-year-old hypertensive man who underwent femoral coronary angiography after a myocardial infarction.

Infective endocarditis caused by Escherichia coli of a native mitral valve.

Escherichia coli is a rare cause of infectious endocarditis. We report a clinical case of E. coli endocarditis of a native mitral valve in a young 26-year-old woman with a recurrent urinary tract infection who had a high fever for one week despite probabilistic treatment with amoxcillin-clavulanic acid 3 g per day. The patient was successfully treated with antibiotics and recovered without surgery.

The occurrence of a multidrug-resistant tuberculous retropharyngeal abscess in an immunocompetent patient: A case report.

Retropharyngeal abscess is an uncommon location of tuberculosis (TB). In this report, we describe a multidrug-resistant tuberculous retropharyngeal abscess in a 21-year-old female patient who was treated for lymph node TB for one year. CT scan revealed a large retropharyngeal abscess that was aspirated intraorally under local anesthesia. The diagnosis of TB was retained by molecular and histological study. GeneXpert MTB/ RIF (Cepheid, Sunnyvale, CA, USA),performed on the pus, showed rifampicin resistance and a first- and second-line drug resistance test using Genotype MTBDRplus VER.2 and MTBDRsl VER.1 (Hain Lifescience GmbH, Nehren, Germany) showed TB highly resistant to rifampicin, isoniazid, and aminoglycosides. Treatment is primarily medical as it combines specific antituberculous antibiotics, and aspiration for drainage of the abscess. Our patient was put on long-term 2nd line anti-TB treatment.

Breast tuberculosis: A forgotten diagnosis.

Breast tuberculosis is a rare entity even in endemic countries. It is often considered as a diagnostic conundrum given the non-specific clinical and imaging characteristics. Therefore, the definitive diagnosis is based on the identification of bacilli by microbiological or histopathological examination. We report the case of a 52-year-old woman, with a 2-month history of a painful lump of the left breast. The imaging features were consistent with a breast abscess. The purulent collection was aspirated and a tuberculous mammary abscess was diagnosed by molecular biology (GeneXpert). Through this observation, we discuss the clinical, radiological and biological signs by highlighting the contribution of molecular biology in the rapid management of this rare pathology.

Whole-Genome Sequencing of Two Moroccan Mycobacterium tuberculosis Strains.

Mycobacterium tuberculosis is known to cause pulmonary and extrapulmonary tuberculosis. In Morocco, the spread of multidrug-resistant (MDR) tuberculosis (TB) has become a major challenge. Here, we announce the draft genome sequences of two Mycobacterium tuberculosis strains, MTB1 and MTB2, isolated from patients with pulmonary tuberculosis in Morocco, to describe variants associated with drug resistance.

Septic arthritis of the ankle: Do not forget Pasteurella pneumotropica.

Pasteurella pneumotropica is an important bacterial pathogen in both animals and humans. Most reported Pasteurella infections in humans involve skin and soft tissues, often after an animal bite, scratch, or lick to an open wound. We report a case of septic arthritis with Pasteurella pneumotropica in a diabetic and cardiopathic patient who was the victim of a rat bite in the street, with a good evolution after medical and surgical treatment.

Serious phlegmonous lesion of the hand following an injury by vegetal thorn: Never forget Pasteurella multocida!

Pasteurella multocida can cause serious infections after dog or cat bite. We report here a rare case of hand infection caused by P. multocida consecutive to an injury by a thorn of the prickly pear. It caused an amputation of the distal phalanx of the thumb in a trisomic patient. It is about a 27-year-old man who was admitted to the hospital with swelling and intense pain of the left hand. He reported a sting by a thorn of prickly pear 15 days before. The patient was admitted to proceed with operative irrigation and debridement. The pus was collected for microbiological examination. Microscopic examination after Gram staining revealed small Gram-negative coccobacilli, associated to polymorphonuclear reaction. Culture have objectivated Pasteurella multocida. The isolated strain was susceptible to betalactamins. Patient was treated with ampicillin. Well-conducted antibiotics and repetitive local cares have not prevented local lesions from progressing to necrosis of the soft parts of the thumb and osteitis of the distal phalanx of the thumb. The patient underwent a necrosectomy and an amputation of the distal phalanx. Ampicillin was replaced by amoxicillin/clavulanic acid and after 15 days, progression was clinically and microbiologically favorable. In the case we report, since the patient does not report any exposure or contact with animals, the thorn prick is the source of infection. It was contaminated from the animal reservoir. Taking into account the monomicrobism of the infection, treatment with aminopenicillins was sufficient. Our propositus came to the hospital 15 days after the inoculation of the bacterium. This duration appears to be very late in relation to the acute character of pasteurellosis. This was probably the main reason why the local infection evolved towards osteoarticular complications. That's why, we should consider Pasteurella multocida in case of infection by inoculation, even in the absence of contact with the animals.

[Saccharomyces cerevisiae invasive infection: The first reported case in Morocco]. / Infection invasive à Saccharomyces cerevisiae : le premier cas rapporté au Maroc.

Saccharomyces cerevisiae is a cosmopolitan yeast, widely used in agro-alimentary and pharmaceutical industry. Its impact in human pathology is rare, but maybe still underestimated compared to the real situation. This yeast is currently considered as an emerging and opportunistic pathogen. Risk factors are immunosuppression and intravascular device carrying. Fungemias are the most frequent clinical forms. We report the first case of S. cerevisiae invasive infection described in Morocco, and to propose a review of the literature cases of S. cerevisiae infections described worldwide. A 77-year-old patient, with no notable medical history, who was hospitalized for a upper gastrointestinal stenosis secondary to impassable metastatic gastric tumor. Its history was marked by the onset of septic shock, with S. cerevisiae in his urine and in his blood, with arguments for confirmation of invasion: the presence of several risk factors in the patient, positive direct microbiological examination, abundant and exclusive culture of S. cerevisiae from clinical samples. Species identification was confirmed by the study of biochemical characteristics of the isolated yeast. Confirmation of S. cerevisiae infection requires a clinical suspicion in patients with risk factors, but also a correct microbiological diagnosis.

Annotated Whole-Genome Shotgun Sequence of Multidrug-Resistant Mycobacterium tuberculosis MTB13_M Isolated from Morocco.

Here, we describe the annotated genome sequence of Mycobacterium tuberculosis MTB13_M. The organism was isolated from a sputum sample in Morocco.

Postoperative mediastinitis due to Candida tropicalis: first reported case in Morocco.

Mediastinitis is a rare and serious nosocomial complication of cardiac surgery. It is estimated at less than 2% of cases of median sternotomy. We report the case of a postoperative mediastinitis due to Candida tropicalis. A 9-month-old baby was operated for complete repair of tetralogy of Fallot in which we isolated C. tropicalis from sternal purulent fluid and blood culture. The child did not survive, despite the initiation of antifungal therapy. The management of this type of infection requires an early diagnosis and an appropriate prolonged treatment, associated with effective preventive measures.