[Usefulness of PCR test for the management of a Mycoplasma pneumoniae outbreak in Bethune Hospital (Pas-de-Calais, France)]. / Importance de la PCR dans la gestion d'une épidémie àMycoplasma pneumoniae au centre hospitalier de Béthune (Pas-de-Calais).

SUBJECT: Molecular amplification (PCR) provides adequate rapid and specific diagnosis of Mycoplasma pneumoniae infection (first agent responsible for community-wide bacterial pneumonia in children above 5 years of age). METHOD: Positive (Chlamylège(®), Argène) PCR in nasopharyngeal aspirate, respiratory samples and nasopharyngeal swab and/or positive serological test (ELISA). RESULTS: Diagnosis of M. pneumoniae infection in 39 cases: 31 between September and December 2008 (30 children and one adult) and eight since June 2009 (three adults and five children). Children (mean age: 3.6years) were hospitalized in 88.6% of cases, mean hospitalization duration was 2.9 days for respiratory tract infections, mainly due to lack of response to ß-lactamines therapy (65.7%). Four adults (mean age: 29.5 years) presented a pneumonia, with hospitalization for three of them with one in intensive care unit. Twenty-eight PCR have proved positive (87%): without associated serology (13), eight negative serologies, IgG and IgM positive (five), and IgG alone (two). Seven patients had only serological test for diagnosis: IgM±IgG. For two children, IgM positive only in isolation, with a PCR probably false negative. CONCLUSION: The sensitivity of the serology in the diagnosis of mycoplasma infection is limited: IgM, which appear traditionally 1 week after clinical signs are mostly inexistent for adults and IgG rise at a later stage. Early diagnosis of child pneumoniae by PCR helped rapidly characterize this epidemic phenomenon and adapt the treatment.

[Implementation of vanA and vanB genes by PCR technique research interest in system (Xpert vanA/vanB CepheidR) closed in a laboratory of microbiology in managing an outbreak to Enterococcus faecium resistant glycopeptide (EfRG)]. / Intérêt de la mise en place de la recherche des gènes vanA et vanB par technique PCR en système clos (Xpert vanA/vanB Cepheid(®)) dans un laboratoire de microbiologie dans le cadre de la gestion d'une épidémie àEnterococcus faecium résistant aux glycopeptides (EfRG).

SUBJECT: The closed system PCR for the rapid detection of vanA and vanB genes (Xpert vanA/vanB Cepheid(®)) was evaluated in our laboratory, to improve the rapidity of the response and thus the management of patients and isolation measures during two GRE outbreaks. METHOD: From March to December2009, 565 samples were analysed by PCR associated to bacterial culture initially for all samples for 2months (n = 75), and thereafter for PCR-positive samples only. RESULTS: In this study, sensitivity and negative predictive values of the PCR were 100%. Specificity was evaluated in the presence and absence of outbreak: 69.3 and 76.8% respectively. The variability of false positive rates between units were lower in nonepidemic than during epidemic phase. The global false positive rate was 23.9%. CONCLUSION: This easy-to-use technology provides rapid results… four samples are tested in 1h versus 72h for culture. Despite its reagent cost, it represents an important hospital diagnostic tool: improvement of the management of cohorting areas and patient transfer between units, adaptation of isolation measures and treatments. However, culture remains necessary to confirm any positive result obtained by PCR and for epidemiological surveillance.

[Outbreak of vancomycin-resistant Enterococcus faecium (Van B) at the Bethune Hospital (France). Two point-prevalence surveys: May 2008 and January 2009]. / Epidémie à Enterococcus faecium résistant à la vancomycine de type Van B au centre hospitalier de Béthune (Pas-de-Calais). Réalisation de deux enquêtes de prévalence en mai 2008 et en janvier 2009.

OBJECTIVE: An outbreak of vancomycin-resistant Enterococcus faecium (E. faecium) occurred in the Bethune Hospital since March 2008 (two consecutive waves). To control this outbreak, two-point prevalence surveys were conducted in May 2008 and January 2009 in inpatients hospitalised more than 24 hours on previously non-affected wards. METHODS: In each ward, information was given to inpatients, administrative and medical data were collected, rectal swabs or stool samples were performed and cultured on chromogenic media. Data were anonymised, and Epidata software was used for the analysis. RESULTS: In May 2008, nine patients were found to be colonized with vancomycin-resistant E. faecium among the 239 patients evaluated (prevalence : 3.76%), and three new wards were affected: neurology ward, general surgical ward, and emergency department observation unit. In January 2009, only one patient, hospitalised in cardiac intensive care unit, was colonised among the 157 patients evaluated (prevalence 0.63%). CONCLUSIONS: These two-point prevalence surveys identified the reservoir of vancomycin-resistant E. faecium carriage, and were thus helpful to contain the two epidemic waves at the Bethune Hospital. A cohorting of the colonised inpatients was performed. Five secondary colonisation cases were detected among the 181 contact patients in May 2008, and no secondary case among 32 contact patients in January 2009.

Specific secretion of gel-forming mucins and TFF peptides in HT-29 cells of mucin-secreting phenotype.

Trefoil factor family (TFF) peptides are typical secretory products of mucin-producing cells, e.g. of the gastrointestinal tract. Here, the expression and secretion of mucins and TFF peptides was studied in the HT-29 cell line throughout cellular growth and differentiation in relation to a mucin-secreting (HT-29 MTX) or an enterocyte-like (HT-29 G(-)) phenotype. mRNAs of several MUC and TFF genes were expressed in both cell subpopulations. However, for most MUC and TFF genes, the expression appeared strongly induced with the differentiation into the mucin-secreting phenotype. On the other hand, TFF2 was specifically expressed in the mucin-secreting HT-29 MTX cells. The differentiation of HT-29 MTX cells into the mucin-secreting phenotype was characterised by secretion of the gel-forming mucins MUC2, MUC5AC, and MUC5B, however, according to a different pattern in the course of differentiation. A significant amount of TFF1 and TFF3 was secreted after differentiation, also according to a different pattern, whereas TFF2 was only faintly detected. Secretagogues, known to induce the secretion of mucus, increased the secretion of all three TFF peptides. In contrast, neither a secretory mucin nor a TFF peptide was found in the culture medium of HT-29 G(-) cells. Overlay assays indicated that HT-29 MTX mucins bound to secretory peptides of HT-29 MTX cells with relative molecular mass similar to TFF peptides. TFF1 and TFF3 were specifically localised in the mucus layer of HT-29 MTX cells by confocal microscopy. Finally, the secretion of TFF peptides and mucins appears as a co-ordinated process which only occurs after differentiation into goblet cell-like phenotype.

[Nocardia otitidiscaviarum, cutaneous infection in a patient receiving long-term corticosteroid treatment]. / Nocardiose cutanée à Nocardia otitidiscaviarum chez un patient traité par corticothérapie au long cours.

A 62-year-old man, under long-term corticosteroid therapy for pigeon breeder's disease, was admitted to endocrinology disease department for cutaneous abscess on back, limbs and scalp. Culture of various bacteriological samples (cutaneous abscess, blood culture) isolated Nocardia otitidiscaviarum. The patient was treated by trimethoprime-sulfametoxazole during several weeks with abscess disappearance. Our laboratory quickly identificatied a bacteria belonging to the Nocardia genus, with simple technique, later confirmed by a specialized laboratory (Pr. Boiron Claude Bernard University Lyon I) with identification of Nocardia otitidiscaviarum. The proof of pulmonary nocardiosis could not be established despite the existente of several risk factors. Prognosis is poor for immunocompromised patients, but the secondary cutaneous dissemination phase presented a favourable evolution under antibiotic therapy.

[Bacteremia and French computerized disease surveillance system: financial valorisation of an infectious diseases specialist in a hospital]. / Bactériémies et Programme de médicalisation des systèmes d'information : valorisation financière de l'infectiologue à l'hôpital.

OBJECTIVE: Bacteremia surveillance is a mission assumed by the referent person for antimicrobial therapy. We propose an original financial valorization of this activity, using the computerized disease surveillance system (CDSS). MATERIAL AND METHODS: A database collecting community-acquired and care-associated bacteremia was created on January 1, 2009 at the Bethune Hospital, France, using EPI-Info software (EPI Data). This database was used to complete missing data (presence of bacteremia, origin [community-acquired or care-associated], site of infection) in CDSS codes of patients hospitalized in surgical and medical wards (410 beds) during 2009. Financial benefit was assessed by the difference of funds allocated on the basis of CDSS, before and after completion of the missing data. RESULTS: In 2009, 383 out of the 35,000 patients presented with bacteremia. When missing CDSS codes were added, a financial gain of 229,291 euros was obtained, concerning 64 patients. CONCLUSION: Bacteremia surveillance is a transversal task based on quality of care, which may have a positive financial impact. This study may be helpful for clinicians with transversal activities, for whom financial valorization is difficult to implement in the CDSS, particularly without hospitalization beds. The lack of complete notification in the CDSS may cause a substantial financial loss.

[Diagnosis of herpetic esophagitis in the immunocompetent subject by PCR (Herpès Consensus Générique-Argène). Report of six cases]. / Diagnostic d'oesophagite herpétique à HSV1 chez le sujet immunocompétent par PCR (Herpès Consensus Générique-Argène). A propos de six cas.

AIM OF THE WORK: We have researched and identified Herpes viruses on the esophageal biopsies taken during the period between September 2006 and March 2008 for 15 suspected patients. PATIENTS AND METHODS: The esophageal biopsies were transferred to the laboratory being conserved in physiological serum and frozen at -80 degrees C for PCR. A fragment was conserved for histopathological analysis. The specimen was defrozen and refrozen in liquid azote (to limit the inhibitors) and crushed to the powder form. Extraction was then done following the prerecognised protocol (Herpès Consensus Générique-"Argene"). That kit allows the amplification consensus of the viral genome of the most frequently encountered Herpes family virus: HSV1, HSV2, CMV, VZV, EBV and HHV6. The identification of the implicated virus was done by the Hybridowell Herpes Identification (Argene) kit in parallel with the migration of SDS gel of the obtained amplifications. RESULTS: HSV1 was identified in seven esophageal biopsies between the 15 studied. HHV6 and the association HHV6/EBV for two patients and only one biopsy had inconclusive. The endoscopy and the histopathological examination had confirmed ulcerated esophagitis with cytopathogene aspect in favour of viral infection for six patients. CONCLUSION: In absence of inhibitors, the adaptation of the extraction technique of the fragments of tissue for few millimetres and the amplifications by PCR had allowed rapid confirmation of the diagnosis of herpetic esophagitis secondary to HSV1 even before the results of the histopathological examination. Treatment by acyclovir entrained regression of the disease.