PubMed Health⌕ Search

Biomedical subjects

V Jarlier

Publications and source records attributed to V Jarlier.

At least 73 records · Page 4Linked to original sources

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↗

[Surveillance of multiresistant bacteria: justification, role of the laboratory, indicators, and recent French data].

In France, several organizations have given priority status to programs aimed at controlling outbreaks of nosocomial infections due to multiresistant bacteria (MRB), most notably methicillin-resistant Staphylococcus aureus (MRSA) and extended-spectrum beta-lactamase-producing Enterobacteriaceae (ESBLE), thus recognizing the widespread distribution and marked pathogenic potential of MRB. To evaluate the impact of preventive measures, surveillance systems have been set up by bacteriology laboratories. These systems rest on a number of indicators: (1) the percentage of strains of a given bacterial species recovered from diagnostic specimens that were multiresistant; (2) the number of new patients (cases) with recovery of a MRB strain from at least one diagnostic specimen (or at least one screening specimen); (3) the incidence, or incidence density, of cases per 1000 hospital days or 100 admissions; (4) the distinction between cases acquired in and transported to the hospital or department. Studies conducted in France between 1990 and 1995 found that the percentage of Klebsiella pneumoniae that were multiresistant was around 20% overall and 25 to 35% among nosocomial strains, and that the prevalence and incidence of cases were about 0.1 to 0.3 per 100 admissions. The proportion of S. aureus strains exhibiting resistance to methicillin was 35 to 40% overall and 50% among nosocomial strains, and the prevalence and incidence of cases ranged from 0.5 to 1 per 100 admissions.

Bacteria↗

Nonradioactive single-strand conformation polymorphism analysis for detection of fluoroquinolone resistance in mycobacteria.

A simple, rapid, and nonradioactive method for routine detection of fluoroquinolone resistance in mycobacteria is described. A single-strand conformation polymorphism (SSCP) methodology, based on the use of mini-gels and silver staining of DNA, was optimized for the analysis of denatured DNA products obtained by polymerase chain reaction (PCR) from the gyrA gene involved in fluoroquinolone resistance in mycobacteria. The method was successfully applied to fluoroquinolone-susceptible and -resistant laboratory strains of Mycobacterium smegmatis and to clinical strains of Mycobacterium tuberculosis isolated from patients who developed resistance during the course of fluoroquinolone treatment.

Anti-Infective Agents↗

[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↗

Antibiotic susceptibility in aerobic gram-negative bacilli isolated in intensive care units in 39 French teaching hospitals (ICU study).

OBJECTIVE: Evaluation of the distribution and antibiotic susceptibility of the aerobic gram-negative bacilli (AGNB) isolated from patients in intensive care units (ICU study). DESIGN AND SETTING: Microbiological study carried out in 1991 in 39 teaching hospitals. A standardized method was used to determine the minimum inhibitory concentrations of 12 antibiotics against 3366 strains of AGNB (close to 100 strains per hospital) during a period of 3 months. RESULTS: The 2773 initial strains (i.e., the first AGNB isolate for a given species and a given patient) were mainly isolated from the respiratory tract (34.4%), urinary tract (23%), or blood (9.6%) and were mainly Pseudomonas aeruginosa (22.9%), Escherichia coli (22%), Acinetobacter (9.7%), and Klebsiella pneumoniae (8.3%). E. coli was prominent in urine and blood and P. aeruginosa in the respiratory tract. Overall, the rate of susceptibility of AGNB was 58 to 65% to piperacillin, cefotaxime, and gentamicin; 69 to 75% to aztreonam, tobramycin, and ciprofloxacin; 83% to ceftazidime; and 91% to imipenem. The overall rates of susceptibility were higher for the initial strains isolated from blood than for those from the urinary or respiratory tracts, mostly reflecting differences in species distribution. Susceptibility rates were lower for the 593 repeat strains (i.e., all the subsequent isolates for a given species and a given patient) than for the initial strains, mostly due to the higher proportion of resistant species (P. aeruginosa 45.9%) but also due to the difference in susceptibility rates for some species-antibiotic combinations. Concomitant resistance (i.e., resistance to several antibiotics due to independent mechanisms of resistance) was marked between beta-lactams and aminoglycosides or quinolones, particularly in P. aeruginosa and K. pneumoniae. CONCLUSIONS: Rates of resistance in AGNB as a whole and in particular species (P. aeruginosa, Klebsiella), as well as frequency of concomitant resistance found in the French ICU study, were higher than those found in ICU studies conducted with the same methodology in Belgium, The Netherlands, and Germany, which may reflect differences in case mix.

Bacteria, Aerobic↗

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↗

Sequence analysis, purification, and study of inhibition by 4-quinolones of the DNA gyrase from Mycobacterium smegmatis.

We determined the nucleotide sequence of a 6-kb DNA region harboring the recF, orf192, gyrB, and gyrA genes from Mycobacterium smegmatis mc(2)155. The amino acid sequences deduced from gyrA and gyrB displayed 89 and 86% identity, respectively, with the DNA gyrase from Mycobacterium tuberculosis, and 67 and 65% identity, respectively, with that from Streptomyces coelicolor. An open reading frame encoding the C-terminal region of the M. smegmatis RecF polypeptide was found upstream from gyrB and was 57% identical to the open reading frame encoding the C-terminal region of the S. coelicolor RecF protein. The gene orf192 was identified between recF and gyrB and was 39% identical to orf191 found in S. coelicolor in the recF-gyrB region. The M. smegmatis DNA gyrase, which was purified by affinity chromatography on novobiocin-Sepharose, consisted of two polypeptides with apparent molecular masses of 98 and 80 kDa. Determination of the N-terminal amino acid sequence of the B subunit confirmed GTG as the start codon in gyrB. Analysis of the supercoiling activity of the enzyme indicated that the M. smegmatis DNA gyrase was characterized by a specific activity equivalent to that of the Escherichia coli DNA gyrase. Inhibition of this activity by 4-quinolones was investigated by determining the 50% inhibitory concentrations (IC50S) of nalidixic acid, ofloxacin, and ciprofloxacin. The results indicated that the inhibitory activities of these drugs against the M. smegmatis DNA gyrase were markedly lower than those previously reported for the E. coli DNA gyrase. The results also suggested that the higher levels of activity of ofloxacin and ciprofloxacin against M. smegmatis (MICs, 0.5 to 1 microgram/ml), in contrast to that of nalidixic acid (MIC, 256 micrograms/ml), could be related to the higher inhibitory activities of fluoroquinolones against the DNA gyrase from this species (IC50S, 7 to 14 micrograms/ml) compared with that of nalidixic acid (IC50, 1,400 micrograms/ml).

4-Quinolones↗

[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↗

Lack of additive effect between mechanisms of resistance to carbapenems and other beta-lactam agents in Pseudomonas aeruginosa.

Eighty-nine clinical isolates resistant (n = 61) or susceptible (n = 28) to imipenem and exhibiting the main patterns of susceptibility to other beta-lactam agents (wild type pattern, penicillinase pattern, constitutive cephalosporinase pattern) were studied in order to investigate (i) the mechanism of resistance involved and (ii) whether resistance to carbapenems affects the level of resistance to other beta-lactam agents and, conversely, if resistance to other beta-lactam agents affects the level of resistance to carbapenems. For this purpose, the presence of OprD protein in the cell wall was detected by Western blot and beta-lactamase activity by spectrophotometric assay and isoelectric focusing. OprD expression was not detectable in the imipenem-resistant (MIC > or = 16 micrograms/ml) strains. It was decreased in half the strains for which MICs of imipenem were 2 to 8 micrograms/ml and was close to a normal level in the most susceptible strains (MIC < or = 1 microgram/ml), thus demonstrating a direct correlation between the level of susceptibility to imipenem and the level of OprD expression. No imipenemase activity was detected in imipenem-resistant strains. Synergy between imipenem or meropenem and BRL 42715 was observed for all of the strains, demonstrating the role of cephalosporinase in carbapenem resistance. Within each pattern of susceptibility, the mean MICs of beta-lactam agents other than carbapenems were similar, whether the strains were susceptible or resistant to imipenem. Conversely, the mean MICs of imipenem or meropenem for either the imipenem-resistant or the imipenem-susceptible strains were similar, regardless of the susceptibility of these strains to the other beta-lactam agents. Thus, when several mechanisms of resistance to beta-lactam agents are present in the same strain of Pseudomonas aeruginosa, there is no additive effect between these mechanisms.

Anti-Bacterial Agents↗

Prognostic significance of bacterial infection in bleeding cirrhotic patients: a prospective study.

BACKGROUND/AIMS: In cirrhotic patients, bacterial infection is frequently associated with gastrointestinal bleeding and seems to increase mortality. The aims of this study were to determine the incidence of bacterial infections in bleeding cirrhotic patients and the influence of infections on the risk of rebleeding and death. METHODS: Cirrhotic patients admitted for gastrointestinal bleeding who had not received antimicrobial chemotherapy in the previous 7 days were included. Blood, urine, and ascitic fluid cultures were systematically performed 1, 2, 4, and 7 days after admission. RESULTS: Sixty-four patients were enrolled. Forty-two bacterial infections were documented in 23 patients (36%) within 7 days of admission. In patients with bacterial infection, mean Child-Pugh score and mean number of blood units transfused were significantly higher, early rebleeding was more frequent (43.5% vs. 9.8%; P < 0.01), and 4-week mortality was higher (47.8% vs. 14.6%; P < 0.01). Multivariate analysis only identified bacterial infections as predictive of early rebleeding (P < 0.02) and a high Child-Pugh score as predictive of death (P < 0.001). CONCLUSIONS: In bleeding cirrhotic patients, bacterial infections only increase the risk of early rebleeding, and mortality is related to the severity of cirrhosis.

Adolescent↗

Acquired resistance in Mycobacterium avium complex strains isolated from AIDS patients and beige mice during treatment with clarithromycin.

Clarithromycin has been reported to select clarithromycin resistant mutants of Mycobacterium avium complex (MAC) during treatment with clarithromycin in AIDS patients and beige mice. We selected resistant mutants in vitro at a frequency of 5 x 10(-9). Clarithromycin resistant strains of MAC isolated in AIDS patients and beige mice as well as derivatives selected in vitro had a unique pattern of acquired cross-resistance to macrolides and related antibiotics. In contrast, the pattern of resistance to non-macrolide antibiotics remained unchanged in clarithromycin resistant strains. A dramatic decrease in ribosome affinity for clarithromycin and erythromycin was found in clarithromycin resistant strains, but no mutation was found in the peptidyl domain of the 23S rRNA, indicating that another ribosomal modification is involved.

Acquired Immunodeficiency Syndrome↗

Role of mutations in DNA gyrase genes in ciprofloxacin resistance of Pseudomonas aeruginosa susceptible or resistant to imipenem.

In Pseudomonas aeruginosa, resistance to imipenem is mainly related to a lack of protein OprD and resistance to fluoroquinolones is mainly related to alterations in DNA gyrase. However, strains cross resistant to fluoroquinolones and imipenem have been selected in vitro and in vivo with fluoroquinolones. We investigated the mechanisms of resistance to fluoroquinolones in 30 clinical strains of P. aeruginosa resistant to ciprofloxacin (mean MIC, >8 micrograms/ml), 20 of which were also resistant to imipenem (mean MIC, >16 micrograms/ml). By immunoblotting, OprD levels were markedly decreased in all of the imipenem-resistant strains. Plasmids carrying the wild-type gyrA gene (pPAW207) or gyrB gene (pPBW801) of Escherichia coli were introduced into each strain by transformation. MICs of imipenem did not change after transformation, whereas those of ciprofloxacin and sparfloxacin dramatically decreased (25- to 70-fold) for all of the strains. For 28 of them (8 susceptible and 20 resistant to imipenem), complementation was obtained with pPAW207 but not with pPBW801. After complementation, the geometric mean MICs of ciprofloxacin and sparfloxacin (MICs of 0.3 microgram/ml and 0.5 microgram/ml, respectively) were as low as those for wild-type strains. Complementation was obtained only with pPBW801 for one strain and with pPAW207 and pPBW801 for one strain highly resistant to fluoroquinolones. These results demonstrate that in clinical practice, gyrA mutations are the major mechanism of resistance to fluoroquinolones even in the strains of P. aeruginosa resistant to imipenem and lacking OprD, concomitant resistance to these drugs being the result of the addition of at least two independent mechanisms.

Anti-Infective Agents↗

Sequences of conserved region in the A subunit of DNA gyrase from nine species of the genus Mycobacterium: phylogenetic analysis and implication for intrinsic susceptibility to quinolones.

The sequences of a conserved region in the A subunit of DNA gyrase corresponding to the quinolone resistance-determining region were determined for nine mycobacterial species and were compared. Although the nucleotide sequences were highly conserved, they clearly differentiated one species from another. The results of the phylogenetic analysis based on the sequences of the quinolone resistance-determining regions were compared with those provided by the 16S rRNA sequences. Deduced amino acid sequences were identical within the nine species except for amino acid 83, which was frequently involved in acquired resistance to quinolones in many genera, including mycobacteria. The presence at position 83 of an alanine for seven mycobacterial species (M. tuberculosis, M. bovis BCG, M. leprae, M. avium, M. kansasii, M. chelonae, and M. smegmatis) and of a serine for the two remaining mycobacterial species (M. fortuitum and M. aurum) correlated well with the MICs of ofloxacin for both groups of species, suggesting the role of this residue in intrinsic susceptibility to quinolones in mycobacteria.

4-Quinolones↗