Proposition of a safe Mycobacterium tuberculosis complex denaturation method that does not compromise the integrity of DNA for whole-genome sequencing.

Whole-genome sequencing plays now a leading role in epidemiologic studies of tuberculosis. DNA extraction of Mycobacterium tuberculosis complex (MTBC) requires complete inactivation of the strains, to be handled for further molecular procedures. In this study we compared two chloroform-based denaturation methods (one with a step of heat killing, one without) to a traditional heat inactivation method. Our results showed that 40% of the strains of MTBC treated by the traditional protocol resulted in a positive culture whereas no culture was observed with the two chloroform-based protocols. The DNA extracts obtained with chloroform-based protocols preparation were successfully used for whole-genome sequencing. We recommend inactivation with our rapid and efficient denaturation method using chloroform without heat killing which met our expectations and biosecurity requirements.

Molecular characterization of Clostridium difficile clinical isolates in a geriatric hospital.

The discriminatory potential of a combination of various typing methods was evaluated on a set of 21 Clostridium difficile isolates obtained from symptomatic patients hospitalized in a geriatric unit and 7 non-toxigenic isolates from the same hospital. Isolates were firstly serotyped and toxinotyped. Of the 28 isolates, 19 belonged to serogroup A. PCR-ribotyping and PCR-RFLP on the fliC and slpA genes were then applied to these 19 isolates. The results suggest that the combination of PCR-ribotyping with PCR-RFLP analysis of slpA could be more discriminatory and suitable for studying C. difficile epidemiology.

Tuberculosis in adolescents: A French retrospective study of 52 cases.

BACKGROUND: The only available data about tuberculosis (TB) among adolescents date back to the 1980s, although the incidence of tuberculosis has been increasing in this age group. METHODS: Medical records were reviewed for all adolescents aged 12 to 18 years hospitalized with the diagnosis of TB in Avicenne/Jean Verdier Teaching hospital (Seine-Saint-Denis, suburb of Paris) between September 2000 and December 2004. RESULTS: Of the 52 patients identified, 52% were female. Median age at diagnosis was 15 years (range, 12-18 years). The proportion of adolescents known to be born abroad was 90%. Diagnoses resulted from the examination of a sick child in 79% of cases, a case contact investigation of an adult suspected of having TB in 19% and routine tuberculin skin test in 2%. Twenty-seven of 52 patients (52%) had isolated pulmonary disease. Sixteen patients (31%) had pulmonary and extrapulmonary TB and 8 cases (17%) had exclusively extrapulmonary disease. The site of extrapulmonary TB included pleural (n = 8), meningitis (n = 4), lymph node (n = 4), peritoneal (n = 5), osteoarticular (n = 3) and genitourinary (n = 1). TB was confirmed by the isolation of Mycobacterium tuberculosis from sputum (n = 21), gastric aspirate (n = 8), bone (n = 1) or cerebrospinal fluid (n = 2). No case had a relapse or recurrence of disease in median 3.2 years of follow up. CONCLUSIONS: Our results indicate that demographic and clinical characteristics of adolescents with TB differed from adults and children. A specific approach to the prevention and treatment of TB in adolescents is absolutely necessary.

Gastrointestinal tuberculosis: 17 cases collected in 4 hospitals in the northeastern suburb of Paris.

UNLABELLED: Gastrointestinal tuberculosis is a rare form of extrapulmonary tuberculosis and its diagnosis can be difficult. AIMS: To analyze the diagnostic and therapeutic characteristics of gastrointestinal tuberculosis. METHODS: Retrospective study from 17 cases collected in 4 hospitals in Seine Saint-Denis between 1987 and 2002. RESULTS: Seventeen cases and 19 localizations were collected: small intestine (N = 7), ileocecum (N = 6), colon (N = 4) and gastroduodenum (N = 2). Two patients had two localizations. Mean age was 43.9 years. Subjects from immigrant populations (76.5%) were preferentially affected. Twenty-three percent of patients (13 tested) were infected by human immunodeficiency virus. Weight-loss and general weakness (88%), abdominal pain (88%), fever (59%), nausea/vomiting (53%) were the predominant symptoms. The delay in diagnosis was 82 days (range: 7-180) and time before specific treatment 31.6 days (range: 7-90). Histological evidence of caseating granuloma was found in six patients. Mycobacterium tuberculosis was detected in six. Digestive imaging was abnormal in 15 patients. Mesenteric lymph nodes were the most common associated site of tuberculosis (N = 8, 47%). Mean duration of treatment was 8.2 months (range: 6-12). Thirteen patients were cured, three died and one was lost to follow up. CONCLUSION: Gastrointestinal tuberculosis is not an uncommon diagnosis in the north-eastern Parisian area, especially among immigrant populations and immunodeficient patients. The most frequent localizations are the small intestine and ileocecum. Diagnosis can be made by pathology and/or bacteriology on endoscopic and/or surgical biopsy samples.

Limited value of angiogenic factors in obese women.

We performed a case-control study assessing the predictive value of sVEGFR and PlGF for preeclampsia in a population with high obesity prevalence. Body mass index and circulating mean concentrations of sVEGFR and PlGF were significantly higher in preeclamptic women. However, a two variable model associating sVEGFR and PlGF dosages was not better than the model including only age and BMI. When the prevalence of obesity is high, the extra benefit of the dosages of angiogenic factors appears to be of limited value.

Prolonged paradoxical response to anti-tuberculous treatment after infliximab.

A 56-year-old woman with ankylosing spondylitis, treated for 3 months with infliximab, developed miliary tuberculosis with mediastinal lymphadenopathies and brain and splenic lesions. After initial improvement under anti-tuberculous therapy, she suffered an unexpectedly prolonged paradoxical worsening with several episodes of lymphadenopathy, including life-threatening ones, over a period of more than 14 months of follow-up. The outcome was favorable as a result of corticosteroid and surgical treatments. This phenomenon reflects a paradoxical reaction precipitated by infliximab withdrawal.

Temporal dynamics of interferon gamma responses in children evaluated for tuberculosis.

BACKGROUND: Development of T-cells based-Interferon gamma (IFNgamma) assays has offered new possibilities for the diagnosis of latent tuberculosis infection (LTBI) and active disease in adults. Few studies have been performed in children, none in France. With reference to the published data on childhood TB epidemiology in the Paris and Ile de France Region, we considered it important to evaluate the performance of IGRA (QuantiFERON TB Gold In Tube(R), QF-TB-IT) in the diagnosis and the follow-up through treatment of LTBI and active TB in a cohort of French children. METHODOLOGY/PRINCIPAL FINDINGS: 131 children were recruited during a prospective and multicentre study (October 2005 and May 2007; Ethical Committee St Louis Hospital, Paris, study number 2005/32). Children were sampled at day 0, 10, 30, 60 (except Healthy Contacts, HC) and 90 for LTBI and HC, and a further day 120, and day 180 for active TB children. Median age was 7.4 years, with 91% of the children BCG vaccinated. LTBI and active TB children undergoing therapy produced significant higher IFNgamma values after 10 days of treatment (p = 0.035). In addition, IFNgamma values were significantly lower at the end of treatment compared to IFNgamma values at day 0, although the number of positive patients was not significantly different between day 0 and end of treatment. CONCLUSIONS/ SIGNIFICANCE: By following quantitative IFNgamma values in each enrolled child with LTBI or active TB and receiving treatment, we were able to detect an increase in the IFNgamma response at day 10 of treatment which might allow the confirmation of a diagnosis. In addition, a decline in IFNgamma values during treatment makes it possible for clinicians to monitor the effect of preventive or curative therapy.

Utilization of 16S ribosomal DNA sequencing for diagnosis of septicemia due to Neisseria elongata subsp. glycolytica in a neutropenic patient.

Septicemia due to Neisseria elongata subsp. glycolytica occurs infrequently. We report a case of septicemia in a patient undergoing antimitotic chemotherapy. Gram-negative coccobacilli were isolated from blood cultures. The identity of the isolate by phenotypic methods was uncertain. In contrast, identity was confirmed by 16S ribosomal DNA sequencing, which appeared to be very useful for correct identification.

Integron-associated antibiotic resistance in Salmonella enterica serovar typhi from Asia.

Eighteen of 25 isolates of Salmonella enterica serovar Typhi were multidrug resistant and contained class 1 integrons with a single cassette, dfrVII or aadA1. The dfrVII-containing integron was likely borne on an IncHI1 plasmid. Salmonella serovar Typhi could become resistant to broad-spectrum cephalosporins by integrating cassettes, such as veb-1, a common cassette in Asia.

[Neuromeningeal tuberculosis in northeastern suburbs of Paris. Nineteen cases]. / La tuberculose neuroméningée dans la banlieue nord-est de Paris.

SUBJECT: Analyze characteristics of neuromeningeal tuberculosis in the northeastern suburbs of Paris. MATERIAL AND METHOD: Retrospective study of 19 observations between 1988 and 1999. RESULTS: Thirteen cases of meningitis, 3 cases of meningitis associated with a tuberculoma and 3 cases with isolated tuberculoma were described. Young subjects (average age 46.4), foreign born (73.7%) were preferentially affected. Three patients were infected by the human immunodeficiency virus. Weight-loss and weakness (84%), fever (79%), headache (63%) dominate the clinical manifestations. The diagnosis delay was 56.6 days, and the therapeutic time, 8.6 days. The search for acid fast bacilli was positive only in one case on direct examination of the cerebrospinal fluid, whereas the culture was positive in 68.7% of the cases. Cerebral imagery was abnormal in13 patients. Average duration of antituberculous treatment was 13,4 months (range: 9-24). Corticosteroids were associated in 12 patients. Thirteen patients were cured, 3 died and 3 were lost to follow up. CONCLUSION: The characteristics of neuromeningeal tuberculosis are not modified, but the diagnosis delay of the neuromeningeal tuberculosis, which conditions the prognosis, remains significant. The optimal duration of the antituberculous treatment and the conditions of the steroid therapy have to be specified.

Anti-phenolic glycolipid-I (PGL-I) determination using blood collection on filter paper in leprosy patients

The authors studied 70 leprosy patients and 20 normal individuals, comparing the traditional sera collection method and the finger prick blood with the conservation on filter paper for specific antibodies against the native phenolic glycolipid-I (PGL-I) from Mycobacterium leprae. The finger prick blood dried on filter paper was eluated in phosphate buffer saline (PBS) containing 0.5 percent gelatin. The classical method for native PGL-I was performed for these eluates, and compared with the antibody determination for sera. It was observed that there is a straight correlation comparing these two methods; although the titles found for the eluates were lower than those obtained for serology. This blood collection method could be useful for investigation of new leprosy cases in field, specially in contacts individuals