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J Grosset

Publications and source records attributed to J Grosset.

At least 55 records · Page 3Linked to original sources

Clinical utility of an amplification test based on ligase chain reaction in pulmonary tuberculosis.

We evaluated the sensitivity and specificity of a new semiautomated direct amplification test (DAT), the LCx-MTB, for the diagnosis of pulmonary tuberculosis (TB) and assessed its positive predictive value by focusing on patients with high clinical and radiologic suspicion of pulmonary TB. Respiratory tract specimens from 32 consecutive patients with high suspicion of active pulmonary TB (case patients) and from 204 control patients were cultured for Mycobacterium tuberculosis and tested by LCx-MTB. Sensitivity and specificity of LCx-MTB when compared with culture was, respectively, 80 and 98%. Pulmonary TB was confirmed in the 32 case patients without knowledge of the LCx results: 18 patients were smear- and culture-positive for M. tuberculosis, and all gave at least one specimen that was LCx-positive. Eight patients were smear-negative culture-positive, and seven gave at least one LCx-positive specimen. LCx-MTB was negative in all the specimens obtained from six patients with smear- and culture-negative TB. A positive LCx-MTB result in a smear negative specimen was 100% predictive that at least one of the case patients' specimens would yield M. tuberculosis. As a consequence, knowledge of the LCx-MTB results at the time of specimen collection could have hastened the start of the antituberculosis therapy in three (21%) smear-negative case patients and could have avoided unnecessary invasive diagnostic procedures in four (29%). We conclude that the sensitivity of LCx-MTB in detecting M. tuberculosis DNA in respiratory tract specimens is similar to other DATs, that LCx-MTB is a reliable test for confirmation of TB in smear-positive patients and that LCx-MTB could be beneficial as a diagnostic step in smear-negative patients with a high suspicion of pulmonary TB.

Antigens, Bacterial↗

Use of DNA fingerprinting for primary surveillance of nosocomial tuberculosis in a large urban hospital: detection of outbreaks in homeless people and migrant workers.

SETTING: A large urban teaching hospital in the southeast of Paris. OBJECTIVE: Primary surveillance of nosocomial transmission of tuberculosis (TB) by systematic restriction fragment length polymorphism analysis (RFLP) of isolates (n = 161) recovered from smear-positive pulmonary TB patients identified from 1 March 1993 to 28 February 1994, and from all TB patients (with any form of tuberculous infection) identified from 1 March 1994 to 30 April 1995. RESULTS: Systematic RFLP analysis revealed 12 clusters of patients (n = 40) infected by strains of Mycobacterium tuberculosis showing matching RFLP patterns. None of the isolates were multidrug-resistant. Compared with non-clustered patients, clustered patients were more likely to be homeless (55% vs 19%, P < or = 0.001), or Africans living in hostels for migrant workers (20% vs 6%, P = 0.01), and had fewer previous admissions to hospital (12% vs 28%, P = 0.05). Further epidemiological investigations showed that the clustered TB cases actually resulted not from nosocomial transmission, but from transmission in the community, very likely in homeless shelters and hostels for migrant workers. CONCLUSION: No nosocomial transmission of TB was identified among the patients included during the study period. Systematic RFLP analysis using hospital-based sampling can detect the spread of TB in specific environments in the community where transmission is occurring.

Adult↗

Characterization of a barley gene coding for an alpha-amylase inhibitor subunit (CMd protein) and analysis of its promoter in transgenic tobacco plants and in maize kernels by microprojectile bombardment.

A gene coding for a barley CMd protein was isolated from a genomic library using a cDNA probe encoding the wheat CM3 protein. Promoter sequence analysis reveals motifs found in genes specifically expressed in endosperm and aleurone cells, as well as TATA and other putative functional boxes. 720 bp of the Hv85.1 CMd protein gene promoter, when fused to a gus coding region, were unable to direct GUS activity in the seeds of transgenic tobacco plants. In contrast, the same construction delivered into immature maize kernels by microprojectile bombardment was able to direct expression of GUS in the outermost cell layers of maize endosperm in both a tissue-specific and a developmentally determined manner.

Amino Acid Sequence↗

Variations of teicoplanin concentrations in neutropenic patients.

The aim of this study was to measure the trough plasma concentrations of teicoplanin in neutropenic patients with a view to optimizing the loading dose. Teicoplanin trough plasma concentrations were followed in 11 neutropenic patients after repeated administration of a 6 mg/kg i.v. bolus. The first three injections were given at 12-h intervals, and the rest every 24 h. Trough plasma concentrations at 48 h varied from 5.6 to 13.1 mg/I. The mean trough plasma concentration-time curve indicated a trend towards accumulation. In conclusion, the loading dose of teicoplanin should be tailored to individual neutropenic patients.

Adolescent↗

Comparison of combination therapy regimens for treatment of human immunodeficiency virus-infected patients with disseminated bacteremia due to Mycobacterium avium. ANRS Trial 033 Curavium Group. Agence Nationale de Recherche sur le Sida.

We conducted a randomized, open-label trial in 42 French hospitals to compare the clinical and bacteriologic efficacy of combination therapy with clarithromycin/clofazimine (Clm/Clof) with that of combination therapy with clarithromycin/rifabutin/ethambutol (Clm/Rib/Eth) as treatment for Mycobacterium avium bacteremia. One hundred forty-four human immunodeficiency virus-seropositive patients older than 18 years of age who had CD4 lymphocyte counts of <100/mm3 and a blood culture positive for M. avium were enrolled in the study. The main measures of outcome were blood cultures, abatement of clinical symptoms (fever), and survival. Treatment success (defined as patient living, either no fever or a reduction of > or = 1 degrees C in initial body temperature, and a blood culture negative for M. avium) was similar in both treatment groups at months 2 and 6. However, following initial resolution of infection, relapse of M. avium bacteremia occurred in more patients in the Clm/Clof group than in the Clm/Rib/Eth group (22 vs. six, respectively; P < .001); these relapses were accompanied by emergence of strains resistant to clarithromycin in 21 and two patients, respectively. In conclusion, combination therapy with Clm/Rib/Eth prevented relapse of mycobacterial disease and, compared with combination therapy with Clm/Clof, was associated with a significant decrease in the emergence of resistant M. avium strains in HIV-infected patients treated for at least 28 weeks.

AIDS-Related Opportunistic Infections↗

Which aminoglycoside or fluoroquinolone is more active against Mycobacterium tuberculosis in mice?

To identify the most active aminoglycoside or fluoroquinolone for the treatment of tuberculosis, the in vivo activities of four different aminoglycosides and three different fluoroquinolones were compared with that of isoniazid (INH) in a murine tuberculosis model. Mice were each inoculated intravenously with 2.3 x 10(7) CFU of Mycobacterium tuberculosis H37Rv. Treatment began the next day (D1) after inoculation and continued for 4 weeks, at the frequency of six times weekly with one of the following regimens: INH, 25 mg/kg; ofloxacin, 200 mg/kg; levofloxacin, 100 or 200 mg/kg; sparfloxacin (SPFX), 50 mg/kg; and streptomycin, kanamycin, amikacin (AMIKA), and isepamicin, all at 200 mg/kg. The dosages of the treatments were presumably equivalent to their clinically tolerated dosages. The severity of infection and effectiveness of the treatment were assessed by the survival rate, spleen weights, gross lung lesions, and the numbers of CFU in the spleens. The results indicate that INH is more bactericidal than any of the aminoglycosides or fluoroquinolones tested, that AMIKA is the most active aminoglycoside, and that SPFX at 50 mg/kg is far more bactericidal than the treatment with other fluoroquinolones.

Aminoglycosides↗

Comparative activities of amikacin against Mycobacterium avium complex in nude and beige mice.

After 4 weeks of treatment, clarithromycin (CLAR) and amikacin showed similar antimicrobial activities against the Mycobacterium avium complex in mice. There was a difference, however, in the effectiveness of the drugs in different types of mice: both drugs displayed bactericidal effects in beige mice but only bacteriostatic effects in nude mice. Because the effectiveness of CLAR is less in nude mice than in beige mice, the predictive value of the nude mouse model for the efficacy of chemotherapy is less than that of the beige mouse model.

Amikacin↗

Prevention of the selection of clarithromycin-resistant Mycobacterium avium-intracellulare complex.

The prevalence of clarithromycin-resistant mutants in untreated bacterial populations of Mycobacterium avium-intracellulare complex (MAC) has been demonstrated to be between 10(-7) and 10(-8) colony-forming units (CFUs) in the beige mouse model. Selection of these mutants occurred during clarithromycin monotherapy if treatment was initiated when the bacterial population size reached approximately 10(8) CFUs per spleen. Likewise, selection of clarithromycin-resistant MAC was induced in AIDS patients during therapy with clarithromycin alone or in combination with drugs that were ineffective for the treatment or prevention of MAC infection. Because the emergence of clarithromycin resistance during preventive therapy was observed exclusively in AIDS patients with CD4+ cell counts < or = 25 cells/microliter, clarithromycin monotherapy can be recommended for the prevention of MAC infection in AIDS patients with CD4+ cell counts of > or = 50 cells/microliter. However, a clarithromycin-containing combination regimen is recommended for patients with CD4+ cell counts < 50 cells/microliter. Since preliminary animal experiments and clinical trials indicate that amikacin, ethambutol or rifabutin in combination with clarithromycin may prevent, or at least delay, the selection of clarithromycin-resistant mutants, further preventive trials are urgently needed to confirm these observations.

AIDS-Related Opportunistic Infections↗

[The bacteriology of tuberculosis and non-tuberculosis mycobacterial infections].

Changing incidence and nature of mycobacterial infections subsequent to the historical regression of tuberculosis and the acquired human immunodeficiency syndrome (AIDS) epidemic, as well as the development of new technical tools for molecular biology, have profoundly modified the methods used for the bacteriological diagnosis of mycobacteria infections. Although microscopic search for acid-fast bacilli, culture and antibiotic resistance tests on Löwenstein-Jensen medium remain the reference methods, more rapid and sophisticated methods are now available. Culture on radiolabeled media using the Bactec system has shortened the delay for positive culture and interpretable antibiotic sensitivity tests. Molecular techniques allow: 1) rapid identification of the most frequently isolated mycobacteria strains, including the most frequent laboratory contaminant M. gordonae, with genome probes; 2) genome typing of M. tuberculosis strains to trace interhuman transmission, detect recurrence or exogenous reinfection or demonstrate laboratory contamination; 3) rapid detection of rifampicin resistance; and 4) direct detection of M. tuberculosis and M. avium in pathological specimens. The role of mycobacteria in the environment causing opportunistic infections, atypical mycobacteria or non-tuberculosis mycobacteria (NTM), particularly the aviaire complex, has grown considerably. Isolation and identification relies on methods used to detect bacilli as well as blood cultures and analysis of fecal matter. NTM are naturally resistant to most of the antituberculosis antibiotics but are sometimes sensitive to aminoglycosides, fluoroquinolones or new macrolides.

AIDS-Related Opportunistic Infections↗

[Cutaneous tuberculosis. A study of 4 cases].

INTRODUCTION: The recent increase in the incidence of tuberculosis has led to the return of cutaneous forms of this disease. In addition, diagnosis can now be made rapidly using genoma amplification. CASE REPORT: Four cases of cutaneous tuberculosis are described in nonimmunosuppressed patients: two cases of lupus vulgaris, including one due to Mycobacterium africanum, and two others of gummas, including one associated with tuberculosis verrucosa. The diagnosis was suggested by epidemiological, clinical, histological and immunological findings and confirmed by culture of the bacilli in 3 cases and by genoma amplification in 1. DISCUSSION: These observations illustrate the difficulties encountered in determining the tuberculosis nature of skin lesions. The clinical presentation, differential diagnosis, the pathophysiology of this disease and the new interest in genoma amplification are discussed.

Adult↗

[Bacteriological diagnosis of tuberculosis].

Ziehl-Neelsen or fluorescence microscopy, culture and drug sensitivity test onto Loewenstein-Jensen medium or with the more rapid radiometric BACTEC system are still the reference diagnostic tests for tuberculosis. However the slow growth of mycobacteria, the severity of the disease caused and the lack of a suitable sub-typing scheme for epidemiological studies have made tuberculosis a popular application for molecular techniques. The four major diagnostic areas molecular techniques have been applied for: rapid identification of cultured mycobacteria; sub-typing of strains by a DNA fingerprinting method in contact tracing; rapid detection of antimicrobial resistance by direct DNA sequencing of target site genes, single strand conformation polymorphism analysis (PCR-SSCP) or line probe assay (LIPA); detection of M. tuberculosis complex in clinical material.

Bacteriological Techniques↗

Nosocomial transmission of tuberculosis among mentally-handicapped patients in a long-term care facility.

SETTING: A long-term care facility at Saint-Brieuc hospital, France. OBJECTIVE: To investigate a nosocomial outbreak of culture-positive pulmonary tuberculosis in 6 (40%) of 15 mentally handicapped HIV-seronegative patients. DESIGN: The factors contributing to the outbreak were analyzed and the restriction fragment length polymorphism (RFLP) patterns of the six Mycobacterium tuberculosis strains were compared. RESULTS: RFLP analysis of the six strains demonstrated an identical banding pattern, thus confirming the spread of a unique strain. A prolonged period of contagiousness due to a delay in diagnosis of the source patient, as well as crowded living conditions in the facility, probably contributed to the outbreak. Surveillance of residents and staff in contact with the source patient resulted in the detection of five secondary cases. Because effective isolation of mentally handicapped patients in the long-term care facility turned out to be difficult, the six case-patients were transferred to the pneumology department, thus limiting the spread of tuberculosis to other residents and staff. CONCLUSIONS: The present outbreak emphasizes the difficulties of implementing control measures for preventing the nosocomial transmission of tuberculosis in long-term care facilities for mentally handicapped patients.

Adult↗

A blind study of the polymerase chain reaction for the detection of Mycobacterium tuberculosis DNA. Azay Mycobacteria Study Group.

SETTING: Nine French laboratories routinely involved in mycobacterial work. OBJECTIVE: To assess the detection of Mycobacterium tuberculosis in experimental samples by polymerase chain reaction (PCR) using the insertion sequence IS6110 as a target for deoxyribonucleic acid (DNA) amplification. DESIGN: Nine laboratories participated in a blind study of the detection of M. tuberculosis by PCR in 20 coded samples containing either a definite number of M. tuberculosis complex (positive samples) or environmental mycobacteria (four samples) or no mycobacteria (five samples). RESULTS: Five laboratories reported false-positive PCR results, with an average rate of 7%. All laboratories except one reported positive PCR results for samples containing 10(5) cfu/ml or more. M. tuberculosis DNA was detected in two thirds of samples containing 10(4) and 10(3) cfu/ml, and in one third of the samples containing 10(2) cfu/ml. CONCLUSION: The results of the study suggest that PCR using IS6110 as a target for DNA amplication is neither very sensitive nor really specific for the detection of M. tuberculosis.

DNA, Bacterial↗

How effective is KRM-1648 in treatment of disseminated Mycobacterium avium complex infections in beige mice?

Although the MICs of 3'-hydroxy-5'-(4-isobutyl-1-piperazinyl)benzoxazinorifamycin, or KRM-1648 (KRM), for Mycobacterium avium complex (MAC) were significantly lower than those of other drugs, its in vivo activity was very weak. Beginning 28 days after inoculation, beige mice that had been infected intravenously with 1.87 x 10(7) CFU of MAC 101 were administered KRM alone, clarithromycin (CLARI) alone, or CLARI plus KRM six times weekly for 16 weeks. In contrast to the mice treated with CLARI-containing regimens, the mortality and the mean spleen weights of mice treated with KRM alone (either 10 or 20 mg/kg of body weight per dose) did not differ significantly from those of untreated mice, their numbers of CFU were very much greater than pretreatment values, and multiplication of MAC was only slightly inhibited. Although monotherapy by KRM selected KRM-resistant mutants, the selection was very weak; the mean number of CFU and the frequency of KRM-resistant mutants increased by no more than 1 order of magnitude after 16 weeks of treatment with KRM at 20 mg/kg per dose. Selection of CLARI-resistant mutants was inhibited but not completely prevented by treatment of the mice with CLARI plus KRM. These results indicate that KRM displayed only a weak bacteriostatic effect against the isolate tested in the beige mouse model; its ability to enhance the antimicrobial effect of CLARI or to prevent emergence of CLARI-resistant mutants was very limited.

Animals↗

Determination of the minimal effective dosages of ofloxacin and sparfloxacin against M. leprae in the mouse foot pad system.

The minimal effective dosages (MEDs) of ofloxacin (OFLO) and sparfloxacin (SPFX) against 10 isolates of Mycobacterium leprae were measured in the mouse foot pad system. The drugs were administered either by gavage or by incorporation into the mouse diet in a range of concentrations. The results demonstrated that the MEDs of OFLO were 4 to 5 times higher than those of SPFX, thus confirming that, on a weight-to-weight basis, the anti-M. leprae activity of SPFX was significantly greater than that of OFLO. The MEDs of OFLO/SPFX measured by gavage were 20 times lower than those measured by incorporating the drug into the mouse diet.

Animals↗

[Analysis of restriction fragment length polymorphism (RFLP) of Mycobacterium tuberculosis strains isolated from patients with several episodes of tuberculosis].

Analysis of restriction fragment length polymorphism (RFLP), using a DNA probe directed against the insertion sequence IS6110, was applied to strains of Mycobacterium tuberculosis successively isolated from four patients. In order to determine the cause of recurrence in these patients, the RFLP patterns of the corresponding isolated were analyzed. The profils obtained from the strains isolated from each of the patients were identical, thus suggesting that a relapse, rather than an exogenous reinfection with a new strain, was the cause of recurrence. The RFLP patterns of successive isolated remained unchanged during periods of time ranging from 5 months to 7 years, and were not modified after development of rifampin resistance. These results demonstrate the stability of the polymorphism detected by the IS6110 probe. Therefore, RFLP analysis is a powerfull epidemiologic tool to distinguish relapse from exogenous reinfection.

AIDS-Related Opportunistic Infections↗

Surveillance of drug resistance for tuberculosis control: why and how?

The resistance of Mycobacterium tuberculosis to antibiotics, which reflects the quality of the chemotherapy applied in the community, is one of the elements of epidemiological surveillance used in national tuberculosis programmes. Measurement of drug resistance poses problems for biologists in standardization of laboratory methods and quality control. The definition of rates of acquired and primary drug resistance also necessitates standardization in the methods used to collect information transmitted by clinicians. Finally, the significance of the rates calculated depends on the choice of the patients sample on which sensitivity tests have been performed. National surveys of drug resistance therefore require multidisciplinary participation in order to select the only useful indicators: rates of primary resistance and of acquired resistance. These indicators, gathered in representative groups of patients over a long period, are a measurement of the impact of modern chemotherapy regimens on bacterial ecology.

Adolescent↗