[Routine use of gene amplification for pertussis diagnosis in children]. / Utilisation en routine de l'amplification génique pour le diagnostic de coqueluche chez l'enfant.

OBJECTIVES: The resurgence of whooping cough observed in France convinced us to develop a specific PCR assay to detect B. pertussis in nasopharyngeal secretions in parallel of the culture. The aim of our study was to show the value of the PCR in routine diagnosis. MATERIAL AND METHODS: From November 1996 to August 2000, in two hospitals located in the Yvelines (France), the children consulting for a cough compatible with the diagnosis of whooping cough were included in this study. A questionnaire including clinical, biological and radiological items was completed for each one of these patients. A culture of Bordetella and a detection by PCR of B. pertussis were carried out on each nasopharyngeal aspirate. The diagnosis of whooping cough was retained if the detection was positive in PCR and/or culture. RESULTS: Among the 215 investigated children with suspected cases of whooping cough, the diagnosis was positive for 45 (20.9%), of which 39 were less than one year old (median: three months). Sixteen (35.5,%) were positive at the same time for both the PCR and the culture, 26 (57.8%) for only PCR and three (6.7%) for only culture. The PCR was positive in 93.3% of the cases. The results were obtained with an average time of 48 hours. The culture was positive in 61.2% of the cases with an average time of six days. The monthly distribution of the cases of whooping cough was very inhomogeneous and of epidemic appearance. The majority of the cases was located between two periods: 42% between November 1996 and September 1997 and 40% between November 1999 and August 2000. Among the infected children, 15 were less than two months old and were not yet vaccinated; among the 24 others infants, a delay in the vaccine calendar was noted in 50% of the cases. Four children between six and 14 years old were correctly vaccinated. The evolution was favourable in all the children. CONCLUSION: The PCR due to its sensitivity, its specificity and its rapidity offers to the clinician a powerful tool for the diagnosis of whooping cough. Nevertheless, the culture must be associated with the PCR, in order to follow the epidemiology and the sensitivity to antibiotics of B. pertussis.

[Outbreak of methicillin-resistant Staphylococcus aureus in general hospital intensive care unit]. / Epidémie à Staphylococcus aureus résistant à la méticilline dans une réanimation polyvalente d'un hôpital général.

Mantes' hospital polyvalent intensive care unit (ICU) experienced an outbreak episode caused by methicillin resistant Staphylococcus aureus (MRSA). Suspicion of physicians was strengthened by observing the weekly reading of multiresistant germs and the significative increase of MRSA carriers incidence rate, compared with the number of admission in the ICU: 5.5% to 11.3%. This outbreak was surprising: it happened immediately after the installation in a new hospital and the reinforcement of nosocomial infection surveillance (systematic screening of every patient admitted to the I.U.C., his isolation if he presents risk factors to multiresistant germs, increasing of handwashing stations). The overlapping period of hospitalisation concerning the 13 patients being reported as SARM carrier, having the same antibiogram, and the epidemic curve suggested a cross contamination. The index case was a MRSA carrier the day of her admission and have had a recent hospitalisation in a high risk unit. MRSA has always been isolated in nasal swab. Six patients among the thirteen carriers developed an infection and have been treated by vancomycin: two systemic infections and four pulmonary infections. The mortality rate was 33% and only one of them seemed to be directly due to MRSA. Area samples were all negative. The clinical staff have been screened with nasal swab. We identified only one nasal MRSA carrier. The pulsed-field gel electrophoresis study showed that 9/11 which have been analysed were identical. This outbreak brought about staff, more sensibilisation to the nosocomial infection and updating of plain hygien rules leaded to its stop five months later.

[Staphylococcus aureus resistant to methicillin. Survey of prevalence in Yvelines hospitals]. / Staphylococcus aureus résistants à la méticilline. Enquête de prévalence dans les hôpitaux des Yvelines.

During 2 months (01.01.95-28.02.95), we noted for each Staphylococcus aureus (SA) isolates, identification and susceptibility methods, hospital size, medical speciality, source, penicillin (P) and methicillin (M) susceptibility. For each non repetitive methicillin resistant SA we noted phenotype of aminoglycoside resistance of SA to 6 antistaphylococci drugs. We isolated 399 SA, SAP sensitives 6.5%, methicillin resistant SA (MRSA) 31.8%. MRSA in acute care unit 28.4%, non acute care unit 61.7%, surgery 28.6%, medicine 36.5%, orthopedics 34.5%, pédiatrics 4%, ICU 38.8%, others 10.3%. MRSA from wound 28.2%, respiratory tract 38.3% blood 22.5%, urine 58.5%, others 25.8% - MRSA kanamycin 1.7%, kanamycin tobramycin 36.2%, kanamycin, tobramycin, gentamicin 56.9% - MRSA erythromycin 79.5%, pefloxacin 91.3%, rifampicin 53.5%, acide fusidique 8.7%, fosfomycine 18.9%, TMP SMZ 5.5%. The proportion MRSA is equivalent among medical specialities predominant in urine cultures and respiratory tract. It should be better to carry on the survey for more than two months in each hospital.

[Nocardia farcinica infection. Cutaneous form in an immunodepressed patient]. / Infection à Nocardia farcinica. Forme cutanée pure chez une malade immunodéprimée.

Nocardiosis is a rare localized or systemic infection caused by bacteria of the Actinomycetaceae family. Nocardia farcinica, recently identified as a distinct species from Nocardia asteroides, characteristically causes severe systemic infections and is particularly resistant to antibiotics. We report a case of nocardiosis observed in a patient receiving general corticosteroid therapy for bullous pemphigoid and who developed a sub-cutaneous abscess of the breast. N. farcinica was identified on puncture specimens and found to be resistant to beta-lactams, aminosides, cyclines, chloramphenicol, fosfomycin and pefloxacin. No dissemination beyond the skin was observed. The abscess was drained and cleaned surgically and cicatrization was uneventful. Six weeks later the patient was again hospitalized for an inflammatory abscess of the left buttocks which was drained surgically. N. farcinica was again identified and a complete work-up eliminated dissemination. Cotrimoxazole was given as a long-term therapy (480 mg trimethoprim, 2.4g sulfamethoxazole) for 6 months and was well tolerated. No recurrence was observed.

[Enterobacteriaceae isolated in nine general hospitals of the Ile-de-France in 1988 and 1989: susceptibility to beta-lactam antibiotics according to the length of hospitalisation]. / Sensibilité aux bêtalactamines en fonction de la durée d'hospitalisation des entérobactéries isolées dans neuf hôpitaux généraux d'Ile-de-France en 1988 et 1989.

Susceptibility to beta-lactam antibiotics of strains of Enterobacteriaceae consecutively isolated in nine general hospitals during a period of 2 months (march and april) has been studied by the disk-agar diffusion method. The separation between susceptible and resistant strains was based on the measure of the inhibition zones centered by 2 disks: cephalothin and ticarcillin. Enterobacteriaceae were divided in 2 groups: strains isolated during the first 48 h of hospitalisation or isolated after. Fifty one per cent of the strains were isolated during the first 48 h: they did not belong to the residential flora of these hospitals. Klebsiella, Proteus indole positive, Providencia, Enterobacter, Serratia were more frequently isolated after 48 h of hospitalisation. Susceptible strains of Klebsiella, Proteus indole positive, Providencia, Serratia were more rarely isolated after 48 h of hospitalisation. E. coli whatever the duration of hospitalisation, is the less frequent susceptible bacterium.

[The value of C-reactive protein in the detection of chorioamnionitis in cases of premature rupture of membranes]. / Valeur de la C-réactive protéine dans le dépistage des chorio-amniotites en cas de rupture prématurée des membranes.

In a context of premature rupture of the membranes, the authors compare the sensitivity and ability to predict the onset of chorio-amnionitis of the conventional clinical signs (hyperthermia, fetal tachycardia, discolored amniotic fluid) and paraclinical signs (hyperleukocytosis and bacteriology of the amniotic fluid) with those of the assay of C reacting protein in the maternal plasma. The latter test is apparently more sensitive and of greater positive predictive value in this disorder.

[Frequency and distribution of beta-lactamases in 1792 strains of Klebsiella pneumoniae in France between 1985 and 1988]. / Fréquence et distribution des bêta-lactamases chez 1,792 souches de Klebsiella pneumoniae isolées en France entre 1985 et 1988.

In october 1985, 1987 and 1988, all the clinical isolates of K. pneumoniae (respectively 530, 654, 590 strains) were collected in 20 hospitals. The beta-lactamases were identified by analytical isoelectrofocusing and by substrate and inhibition profiles. 76 to 81% of the strains produced only one beta-lactamase: SHV-1 type, pI 7.7 (61 to 65%) or PI 7.1 (14%). The TEM-1 betalactamase (pI 5.4) was produced in 1985 by 21% of the strains, 9% in 1987, and 11% in 1988: TEM-2, pI 5.6 by 2% in 1985-87-88. The extended broad spectrum beta-lactamases, able to hydrolyse amino-thiazol-oximino-beta-lactam antibiotics, TEM or SHV type enzymes (SHV-2, pI 7.7, SHV-3, pI 7.1; SHV-4/CAZ-5, pI 7.8; SHV-5/CAZ-4 pI 8.2; CTX-1/TEM-3, pI 6.3) were also detected: 0.75% of the strain (3 strains) in 1985, 8.4% (55 strains) in 1987, 11% (65 strains) in 1988. These extended broad spectrum beta-lactamases were found in 2 hospitals in 1985, 10 in 1987 and 9 in 1988.

[Sensitivity of Pseudomonas aeruginosa and Klebsiella spp. to ceftazidime. Current status in France]. / Sensibilité des Pseudomonas aeruginosa et des Klebsiella à la ceftazidime. Etat actuel en France.

Ceftazidime was tested against 2,224 strains of Pseudomonas aeruginosa obtained from 17 hospitals in April, May and June, 1986 and against 607 strains of Klebsiella pneumoniae and 234 strains of K. oxytoca obtained from 16 hospitals in October, 1987. The MIC's of ceftazidime against P. aeruginosa were distributed normally, with an MIC50 of 2 mg/l and an MIC90 of 4 mg/l. Depending on critical concentrations, 80 per cent of strains were sensitive, 11.4 per cent were of intermediate sensitivity and 0.54 per cent were resistant. There were few differences in results between hospitals. Ninety-two per cent of resistant strains and 45 per cent of intermediate strains (as opposed to 6 per cent of all strains) produced a high-level constitutive cephalosporinase with little variations between centres. The MIC's of ceftazidime against K. pneumoniae and K. oxytoca had a bimodal distribution: 91 per cent of strains were sensitive to 0.25 mg/l, 6 per cent of strains showed intermediate sensitivity and 3 per cent were resistant. All intermediate and resistant strains produced a very broad spectrum beta-lactamase which hydrolyzed some of the third generation cephalosporins: K. pneumoniae 36 CTX-1, 5 SHV-2, and 14 strains producing a recently identified beta-lactamase "CAZ-5/SHV-4"; K. oxytoca 3 CTX-1. These strains were isolated in 10 of the 16 hospitals which took part in the 1987 study. Comparison of these results with those of studies performed in 1984 and 1985 showed a moderate increase in the number of intermediate sensitivity strains of P. aeruginosa and the occasional occurrence, of the epidemic type, in some hospitals of Klebsiella spp. producing very broad spectrum beta-lactamases which were rare in 1985.

Neonatal meningitis due to Gardnerella vaginalis.

A case of meningitis due to Gardnerella vaginalis occurred in a five-day-old newborn who had clinical signs of fever, polypnea and a grey complexion. After treatment with ampicillin, cefotaxime and netilmicin, the patient's condition improved, and no sequelae were observed. The bacterium isolated from a pure culture of the cerebrospinal fluid was identified by biochemical characteristics to be Gardnerella vaginalis, but it was not possible to define the source and mode of contamination.

[In vitro activity of penicillin G and amoxicillin against 14 strains of Streptococcus milleri]. / Activité in vitro de la pénicilline G et de l'amoxicilline vis-à-vis de quatorze souches de Streptococcus milleri.

The MIC/MBC of penicillin G and amoxicillin were studied for 14 strains of Streptococcus milleri from varied specimens of hospitalized patients by broth dilution method. The MIC of penicillin G are between 0.015 and 0.12 mg/l, MIC of amoxicillin between 0.015 and 0.5 mg/l. The MBC of penicillin G are between 0.015 and 0.12 mg/l, MBC of amoxicillin between 0.12 and 0.5 mg/l. MBC/MIC of penicillin G are between 1 and 8. MBC/MIC of amoxicillin are between 1 and 16. The bactericidal effect was studied by killing curves on 5 strains at 2, 4, 6 and 18 hours at different concentrations of penicillin G (0.1, 0.2, 1 mg/l) and of amoxicillin (0.2, 0.5, 1 mg/l). For 3 strains we observed a bactericidal effect with 99.9% or 99.99% of killing between the 6th and 18th hour without difference between penicillin G and amoxicillin, the two further strains were killed respectively by 0.1 mg/l of penicillin G and 0.5 mg/l of amoxicillin and by 0.2 mg/l of penicillin G and 0.5 mg/l of amoxicillin.

[Bactericidal speed of ceftriaxone and cefotaxime on 18 strains of Haemophilus influenzae]. / Vitesse de bactéricidie de la ceftriaxone et du céfotaxime sur dix-huit souches d'Haemophilus influenzae.

The bactericidal effect of ceftriaxone and cefotaxime was studied by killing curves on 18 Haemophilus influenzae at 2, 4, 6 and 24 h. The MIC of ceftriaxone are between 0.0012 and 0.015 mg/l. The MIC of cefotaxime are between 0.006 and 0.03 mg/l. The MBC of ceftriaxone are between 0.0012 and 0.03 mg/l. The MBC of cefotaxime are between 0.006 and 0.03 mg/l. The bactericidal effect was studied at the concentration of 0.06 mg/l; at this concentration we obtained a reduction of 4 log 10 for 3 strains in 6 hours by both drugs. In 24 h, 16 stains give a 99.99% of killing for ceftriaxone and 18 strains for cefotaxime. Our results do not show a significant difference in the bactericidal effect of the two drugs on Haemophilus.

[Study of ampicillin sensitivity of 148 strains of Haemophilus influenzae isolated in 2 general hospitals]. / Etude de la sensibilité à l'ampicilline de 148 souches d'Haemophilus influenzae isolées dans deux hôpitaux généraux.

During one year, by disk diffusion, we have studied in two general hospitals, the susceptibility to ampicillin of Haemophilus influenzae isolated from various clinical specimens, CSF (2 strains), blood (1 strain), ears (6 strains), eyes (18 strains), lower respiratory tract specimens (111 strains), genital specimens (7 strains), and various other specimens (3 strains). The beta-lactamase production was researched systematically with a chromogenic test. At all 148 strains of H. influenzae were examined. Drug resistance was observed for 18 strains (12.2%). All strains produced a beta-lactamase. The study of drug resistance of H. influenzae with the regard of the age showed an higher percentage of resistant strains isolated from children than the percentage of resistant strains isolated from adults. From children, 10 out of 50 strains (20%) were resistant to ampicillin, on the other hand 8 out of 98 (8.2%) strains isolated from adults were resistant to ampicillin.

[Study of the sensitivity to pipemidic acid, pefloxacin and norfloxacin of 444 bacterial strains isolated at a general hospital]. / Etude de la sensibilité à l'acide pipémidique, à la péfloxacine et à la norfloxacine de 444 souches bactériennes isolées dans un hôpital général.

After one year of use of pefloxacin in the intensive care unit, medicine unit and surgery unit and the following recent commercialization without prescription of norfloxacin, we studied for a period of two months (april and may 1986) the susceptibility to pipemidic acid, to pefloxacin and norfloxacin of 444 bacteria strains isolated obtained from clinical specimens. The antibiotic susceptibility is determined by disk diffusion test. We obtained the following results: susceptible methicillin Staphylococcus aureus (SMSA 63 strains): 85.7% PEFS, 11.1% PEFI, 79.4% NORS, 15.9% NORI, resistant methicillin Staphylococcus aureus (RMSA 36 strains): 33.3% PEFS, 2.9% PEFI, 41.6% NORS, 0% NORI; Escherichia coli, Proteus mirabilis, Klebsiella pneumoniae (252 strains): 94% PIPS, 94.8% PEFS, 97.2% NORS; Enterobacter, Proteus indol (+), Providencia and Citrobacter (33 strains): 72.7% PIPS, 72.7% PEFS, 84.8% NORS; Pseudomonas aeruginosa (53 strains): 32% PEFS, 47.2% PEFI, 90.6% NORS, 5.6% NORI; Acinetobacter (7 strains): 57.2% PEFS, 42.8% PEFI, 42.8% NORS, 28.6% NORI. Norfloxacin in active in vitro against the majority of the P. aeruginosa isolated. We found an important methicillin and pefloxacin resistance among the Staphylococcus aureus isolated: 36 strains RMSA (36.4% out of Staphylococcus aureus) and 23 strains RMSA PEFR (63.8% out of RMSA). These later strains were isolated in eight different units mainly in visceral surgery unit and geriatric units but not in intensive care unit. After an epidemiologic study and the following recommendations to physicians and nurses: treatment by pefloxacin should be used only after bacteriological results and associated with an other antibiotic, the handwashing should be frequent and regular to prevent the spread of infection, four months later we isolated 26 strains of RMSA with 7 RMSA PEF (26.9%).2$