clinical strains and transformants had bands of β-lactamase activity with an alkaline pI of 7.6 and 5.4. Polymerase chain reaction (PCR) amplification of the 26 clinical isolates was positive for blaCTX-M and blaTEM (7). The 26 strains of E. coli had the same profile by repetitive-element PCR and pulsed-field gel electrophoresis, while unrelated control strains had very different profiles. Sequencing in strains isolated from four of the patients identified a CTX-M-15 β-lactamase and a TEM-1 β-lactamase. The four strains were related to the phylogenetic group B2 and produced the iutA (ferric aerobactin receptor), YuA (Yersinia siderophore receptor), and fimH (type I fimbriae) virulence factors (8). Incidence of colonization or infection by the culprit strain was 34.3% (12 of 35 patients) within the initial 4-month period and 55.3% (26 of 47 patients) over a 1-year period. Intensified hygienic procedures implemented in January 2002 contributed to a decrease in the number of cases in February only; since then, a regular increase of new cases extended the outbreak and caused problems with controlling it. All urinary tract infections were successfully treated with a 15-day course of trimethoprimsulfamethoxazole; however, reinfection occurred in some. Neither incontinence (p = 0.35), dementia (p = 0.22), nor previous antibiotic treatment (amoxicillin, amoxicillin-clavulanic acid, extended-spectrum cephalosporins, and fluoroquinolones [p = 1.00, 0.30, 0.12, 0.52, respectively]) appeared to be risk factors for infection or colonization in our study, but the number of patients is too small to reach a conclusion. However, patients that were infected or colonized had greater functional impairment, especially incontinence and dementia. Nonambulatory status, decubitus ulcers, and feeding tubes were not risk factors for acquiring ESBL-producing E. coli in our study. The outbreak has not been controlled: 13 patients have persistent digestive-tract colonization. Difficulties encountered in controlling such outbreaks may be explained by several factors. Patients cannot be easily isolated in long-term care facilities. Strict isolation and limitation of activity and mobility cannot always be applied because of their impact on social activities.

Objectives: To correlate human herpesvirus 6 (HHV-6) viral load with pathologic features in graft acute hepatitis of unknown origin.Methods: Liver frozen samples from 26 patients with graft hepatitis of unknown origin were available for HHV-6 DNA quantification.Results: In 10 (38.5%) of 26 liver samples, HHV-6 DNA was detectable, with a median viral load of 3.84 log(10) copies/10(6) cells. Confluent periportal necrosis was observed in 4 of 10 patients and associated with high viral load. These 4 patients responded to antiviral therapy. Mild unspecific hepatitis was observed in 4 patients with low intragraft viral load and in 2 patients with high viral load in a context of deep immunosuppression. Patients with HHV-6 negative graft hepatitis disclosed lobular necrotico-inflammatory activity without periportal necrosis.Conclusions: Our study provides data supporting the pathogenic role of HHV-6 for liver allografts. The presence of confluent periportal necrosis could be a clue for prompt diagnosis of HHV-6 induced graft hepatitis.

The aims of the study were to assess the risk of HHV8 transmission resulting from organ transplantation, and related morbidity in liver, heart and kidney transplant recipients. Donor and recipient serologies were screened between January 1, 2004 and January 1, 2005 using HHV8 indirect immunofluorescence latent assay (latent IFA) and indirect immunofluorescent lytic assay (lytic IFA). Recipients negative for latent IFA with a donor positive for at least one test were sequentially monitored for HHV8 viremia and underwent serological tests over a period of 2 years. The results showed that among 2354 donors, HHV8 seroprevalence was 9.9% (lytic IFA) and 4.4% (latent IFA). A total of 454 organ recipients (281 renal, 116 liver and 57 heart) were monitored over a 2-year period. Seroconversion was observed in 12 patients (cumulative incidence 28%) whose donor had positive latent IFA and in 36 patients (cumulative incidence 29%) whose donors were positive only for lytic IFA, without differences across types of transplants. Positive HHV8 viremia was detected in only 4 out of 89 liver transplant recipients during follow-up and not in recipients of other types of transplant. Two liver transplant recipients and one kidney transplant recipient developed KS. In conclusion, although HHV8 transmission is a frequent event after organ transplantation, HHV8-related morbidity is rather rare but can be life threatening. Donor screening is advisable for monitoring HHV8 seronegative liver transplant recipients.

ABSTRACTA multicentric clinical study was conducted on representative sera from 1,738 European and U.S. subjects for the evaluation of new anti-hepatitis A virus enzyme immunoassays from Bio-Rad Laboratories. Comparison with reference DiaSorin S.p.A. tests confirmed the good performance of Bio-Rad assays (99.85% and 99.47% overall agreement in detecting total antibodies and IgM, respectively).

M et al. Rapid detection of the O25b-ST131 clone of Escherichia coli encompassing the CTX-M-15-producing strains. 2 Clermont O, Lavollay M, Vimont S et al. The CTX-M-15-producing Escherichia coli diffusing clone belongs to a highly virulent B2 phylogenetic subgroup. 3 Karisik E, Ellington MJ, Livermore DM et al. Virulence factors in Escherichia coli with CTX-M-15 and other extended-spectrum b-lactamases in the UK. 4 Deschamps C, Clermont O, Hipeaux MC et al. Multiple acquisitions of CTX-M plasmids in the rare D 2 genotype of Escherichia coli provide evidence for convergent evolution. 7 Ender PT, Gajanana D, Johnston B et al. Transmission of an extended-spectrum-b-lactamase-producing Escherichia coli (sequence type ST131) strain between a father and daughter resulting in septic shock and emphysematous pyelonephritis. A et al. Dissemination of clonally related Escherichia coli strains expressing extended-spectrum b-lactamase CTX-M-15. Sir, The emergence and dissemination of Klebsiella pneumoniae strains harbouring carbapenemases is a serious concern. Klebsiella pneu-moniae carbapenemase (KPC) enzymes belong to molecular class A and are able to hydrolyse most b-lactams including carbapenems. Since the initial report of a KPC b-lactamase from a strain of K. pneu-moniae in 1996, KPC producers have been reported from various geographical regions. Current reports indicate that KPC-producing isolates are widespread in China, Israel, Greece, South America and the USA, where the epidemiology of KPC in the hospital setting is changing. 1 Fortunately, these strains are still rare in western and northern Europe, but their detection remains difficult. 2 Since 2003, patients hospitalized in the surgical ward of our hospital have been systematically screened on admission and weekly thereafter for intestinal carriage of bacteria producing extended-spectrum b-lactamases (ESBLs) and carbapenemases by plating rectal swabs on Drigalski agar containing 0.5 mg/L cefotaxime and MacConkey agar containing 2 mg/L ceftazidime (AES Labora-toire, Combourg, France). We report here four patients with K. pneu-moniae producing KPC-2 and SHV-12. The first case was a patient transferred in July 2009 from Crete for treatment of recurrent angiocholitis on a biliary stent. The patient was negative on the day of admission, but 3 days later a further stool sample grew with ESBL-producing K. pneumoniae. This first isolate was not suspected to produce a carbapenemase since testing for susceptibility to imipenem using a disc diffusion method showed a diameter of 24 mm and an MIC of 1.5 mg/L by Etest (Bio-Rad, Marne la Coquette, France), both considered as susceptible according to the national recommendations of the Antibiogram …

References 1 Novick RP. Autoinduction and signal transduction in the regulation of staphylococcal virulence. Mol Microbiol 2003; 48: 1429–49. 2 Kuroda M, Kuroda H, Oshima T et al. Two-component system VraSR positively modulates the regulation of cell-wall biosynthesis pathway in Staphylococcus aureus. Mol Microbiol 2003; 49: 807–21. 3 Meehl M, Herbert S, Gotz F et al. Interaction of the GraRS two-component system with the VraFG ABC transporter to support vancomycin-intermediate resistance in Staphylococcus aureus. Antimicrob Agents Chemother 2007; 51: 2679–89. 4 Komatsuzawa H, Fujiwara T, Nishi H et al. The gate controlling cell wall synthesis in Staphylococcus aureus. Mol Microbiol 2004; 53: 1221–31. 5 Nishi H, Komatsuzawa H, Fujiwara T et al. Reduced content of lysyl-phosphatidylglycerol in the cytoplasmic membrane affects susceptibility to moenomycin, as well as vancomycin, gentamicin, and antimicrobial peptides, in Staphylococcus aureus. Antimicrob Agents Chemother 2004; 48: 4800–7. 6 Berger-Bachi B, Rohrer S. Factors influencing methicillin resistance in staphylococci. Arch Microbiol 2002; 178: 165–71. J Antimicrob Chemother 2010 doi:10.1093/jac/dkq155 Advance publication 13 May 2010

There is little information on the diverse infectious causes of jaundice and hepatitis in the Asiatic tropics. Serology (hepatitis A, B, C and E, leptospirosis, dengue, rickettsia), antigen tests (dengue), PCR assays (hepatitis A, C and E) and blood cultures (septicaemia) were performed on samples from 392 patients admitted with jaundice or raised transaminases (> or =x3) to Mahosot Hospital, Vientiane, Laos over 3 years. Conservative definitions suggested diagnoses of dengue (8.4%), rickettsioses (7.3%), leptospirosis (6.8%), hepatitis B (4.9%), hepatitis C (4.9%), community-acquired septicaemia (3.3%) and hepatitis E (1.6%). Although anti-hepatitis A virus (HAV) IgM antibody results suggested that 35.8% of patients had acute HAV infections, anti-HAV IgG antibody avidity and HAV PCR suggested that 82% had polyclonal activation and not acute HAV infections. Scrub typhus, murine typhus or leptospirosis were present in 12.8% of patients and were associated with meningism and relatively low AST and ALT elevation. These patients would be expected to respond to empirical doxycycline therapy which, in the absence of virological diagnosis and treatment, may be an appropriate cost-effective intervention in Lao patients with jaundice/hepatitis.

The diagnosis of hepatitis A virus (HAV) infection is based on the detection of anti-HAV IgM. Shortcomings of this serological approach include the persistence of IgM after normalization of liver enzymes or its detection during polyclonal activation of the immune system due to unrelated viral infection or autoimmune diseases. Molecular diagnosis of HAV along with anti-HAV IgG avidity measurement are helpful in case of positive IgM where laboratory evidence of acute hepatitis is absent and there is no epidemiologic link to other cases. Molecular epidemiology allows us to determine whether viruses from different locations are related to each other and provides further understanding of viral epidemiology by identifying sources and transmission modes. It has been demonstrated that the rapid turnover of HAV strains in low-endemicity countries is caused by their introduction by travelers, Growing sequence databases allow for the identification of geographic origin of viral strains. Collaboration between surveillance laboratories, including database sharing, should be promoted for deeper investigation of outbreaks and improved prevention approaches.

We report the successful control of an outbreak caused by imipenem-resistant VIM-1-producing Klebsiella pneumoniae (IR-Kp) in France. This outbreak occurred in a care centre for abdominal surgery that includes a 15-bed liver intensive care unit and performs more than 130 liver transplantations per year. The index case was a patient with acute liver failure transferred from a hospital in Greece for urgent liver transplantation who was carrying IR-Kp at admission as revealed by routine culture of a rectal swab. Infection control measures were undertaken and included contact isolation and promotion of hand hygiene with alcohol-based hand rub solution. Nevertheless, secondary IR-Kp cases were identified during the six following months from 3 December 2003 to 2 June 2004. From 2 June to 21 October, extended infection control measures were set up, such as cohorting IR-Kp carriers, contact patients and new patients in distinct sections with dedicated staff, limiting ward admission, and strict control of patient transfer. They led to a rapid control of the outbreak. The global attack rate of the IR-Kp outbreak was 2.5%, 13% in liver transplant patients and 0.4% in the other patients in the care centre (p<0.005). Systematic screening for IR-Kp of all patients admitted to the care centre is still maintained to date and no secondary IR-Kp case has been detected since 2 June 2004.

Mutations in the internal ribosome entry site (IRES) of hepatitis A virus (HAV) have been associated with enhanced in vitro replication and viral attenuation in animal models. To address the possible role of IRES variability in clinical presentation, IRES sequences were obtained from HAV isolates associated with benign (n = 8) or severe (n = 4) hepatitis. IRES activity was assessed using a bicistronic dual-luciferase expression system in adenocarcinoma (HeLa) and hepatoma (HuH7) cell lines. Activity was higher in HuH7 than in HeLa cells, except for an infrequently isolated genotype IIA strain. Though globally low, significant variation in IRES-dependent translation efficiency was observed between field isolates, reflecting the low but significant genetic variability of this region (94.2% +/- 0.5% nucleotide identity). No mutation was exclusive of benign or severe hepatitis, and variations in IRES activity were not associated with a clinical phenotype, indirectly supporting the preponderance of host factors in determining the clinical presentation.

Purpose of the study. - Very few is known on genotype II hepatitis A virus (HAV) since it is rarely isolated. From 2002 to 2007, the French observatory of HAV identified six sub-genotype IIA strains of which one from a patient having travelled to West Africa. To investigate the possible African origin of sub-genotype IIA, we determined its prevalence among French travellers in 2008 and characterised its genetic variability.Patients and methods. - The 2008 mandatory notification records were screened for travel to Africa. Viral genotype was determined on the nucleotide sequencing of the VP1/2A junction region. The P1 region coding for capsid proteins was used to compare the genetic diversity of IIA isolates to those of other genotypes.Results. - In 2008, five out of 54 patients returning from West Africa were infected by IIA strains and an additional "autochthonous" case was identified. Two more African cases were identified in 2009. A total of 14 IIA isolates (eight African and six "autochthonous") were analysed. Nucleotide and amino-acid variability of IIA sequences was lower than that of the other genotypes. Phylogenetic analysis revealed the clustering of two "autochthonous" cases with African isolates whereas the other ones belonged to a different lineage.Conclusion. - Most IIA strains isolated in France are imported by travellers returning from West Africa. However, the unexplained contamination mode of some "autochthonous" cases suggests another geographical origin to discover or a French reservoir to explore. (C) 2010 Elsevier Masson SAS. All rights reserved.

ABSTRACT Three hepatitis A virus (HAV) genotypes, I, II, and III, divided into subtypes A and B, infect humans. Genotype I is the most frequently reported, while genotype II is hardly ever isolated, and its genetic diversity is unknown. From 2002 to 2007, a French epidemiological survey of HAV identified 6 IIA isolates, mostly from patients who did not travel abroad. The possible African origin of IIA strains was investigated by screening the 2008 mandatory notification records of HAV infection: 171 HAV strains from travelers to West Africa and Morocco were identified. Genotyping was performed by sequencing of the VP1/2A junction in 68 available sera. Entire P1 and 5′ untranslated regions of IIA strains were compared to reference sequences of other genotypes. The screening retrieved 5 imported IIA isolates. An additional autochthonous case and 2 more African cases were identified in 2008 and 2009, respectively. A total of 14 IIA isolates (8 African and 6 autochthonous) were analyzed. IIA sequences presented lower nucleotide and amino acid variability than other genotypes. The highest variability was observed in the N-terminal region of VP1, while for other genotypes the highest variability was observed at the VP1/2A junction. Phylogenetic analysis identified 2 clusters, one gathering all African and two autochthonous cases and a second including only autochthonous isolates. In conclusion, most IIA strains isolated in France are imported by travelers returning from West Africa. However, the unexplained contamination mode of autochthonous cases suggests another, still to be discovered geographical origin or a French reservoir to be explored.

POSTERSquartile of LDLR mRNA expression in the PI biopsy survived with a significantly (Log-Rank test P < 0.05) better survival than those in the three lowest quartiles.Surviving graft (n = 34, median followup 44.8 months, range 8.6-72.3)had significantly higher LDLR (P < 0.01) and NPC1L1 (P < 0.05) mRNA expression in the PI biopsy and HMGCR (P < 0.05) mRNA expression in the PR biopsy than lost grafts (n = 13, median follow-up 6.0 months, range 0.03-43.7).Conclusions: In the settings of human liver transplantation: 1. hepatic LDLR mRNA is overexpressed immediately after graft reperfusion, suggesting an increased hepatocyte cholesterol uptake from blood; 2. hepatic upregulation of genes involved in cholesterol recruitment and synthesis is inversely related to the severity of IRI and is associated with better graft survival.

Since September 2008, 26 cases of hepatitis A with a history of travel to Egypt have been reported in France. Investigations indicate that a common source of contamination linked to Nile river cruises is the most likely explanation of the increase in the number of cases reported in France as well as in several other European Union countries.

Background/Aims: Liver transplantation (LT)in immune-suppressed human immunodeficiency virus(HIV) and hepatitis C virus (HCV) coinfected patients is feasible but raises questions regarding the severity of HCV recurrence on the liver graft and preservation of immune function. We investigated whether LT is deleterious to the immune system.Methods: Fourteen HIV-HCV coinfected patients (HIV viral load [VL] <50 copies/ml; median CD4 count of 276/mm(3) pretransplantation) were grafted for HCV-cirrhosis and followed over 2 years. Nine patients received anti-HCV therapy post-transplantation. HCV and HIV VLs and degree of acute and chronic hepatitis were monitored. Peripheral blood T-cell phenotypes and interferon-gamma (IFN-gamma) immune responses against opportunistic pathogens, HCV, and HIV-1 p24 were evaluated.Results: Median HCV VLs, CD4 counts, T-cell subsets, and IFN-gamma-producing T-cell frequencies against opportunistic pathogens and HIV-1 p24 did not change over time. HCV-specific T cells were observed ex vivo in two patients pretransplantation and in two others post-transplantation. HCV-specific in vitro amplification enabled the detection of HCV-specific IFN-gamma-producing responses in three further patients post-transplantation. Anti-HCV responses were observed independently of anti-HCV therapy and were undetectable in patients with severe hepatitis or liver fibrosis.Conclusions: These results demonstrate that LT in HIV-HCV coinfected patients is not deleterious to the immune system and does not alter immune responses directed against HCV, HIV, or opportunistic pathogens. (C) 2009 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.

The impact of preexisting or acquired Kaposi sarcoma herpesvirus (KSHV) infection in kidney transplant recipients was evaluated in a prospective study. Serum collected from kidney donors and recipients before transplantation were tested for antibodies against KSHV latent nuclear antigen. Three groups of recipients were defined: group A (KSHV+), group B (KSHV-, KSHV+ donor) and group C (donor and recipient KSHV-). Blood was collected from recipients, every 3 months for 3 years, for KSHV viremia (groups A and B), quantitative (group A) and qualitative serology (group B). Data of group C recipients were extracted from a French database. The prevalence of KSHV antibodies was 1.1% in donors and 3.2% in recipients. There were respectively 161, 64 and 4744 recipients in groups A, B and C. In group A, 13% developed Kaposi's sarcoma (KS). Age >53.5 years (p = 0.025) and black skin (p = 0.0054) were associated with KS development. In group B, three recipients developed clinical manifestations related to KSHV infection. There was no difference in terms of survival and graft loss between the three groups. In conclusion, although kidney recipients should be aware of the additional risk of KSHV morbidity, KSHV+ recipients should not be systematically excluded from kidney transplantation.

Following the notification of nine hepatitis A cases clustered in the Cotes d’Armor district in northwestern France, epidemiological, environmental and microbiological investigations were set up in order to identify the source and vehicle of contamination and implement control measures. In total, 111 cases were identified in the outbreak, all of whom lived or had stayed as tourists in the Cotes d’Armor district. Of the cases, 87% had eaten raw shellfish, and 81% specifically oysters. Traceback investigations carried out on raw shellfish consumed by the cases showed that the raw shellfish originated from a single shellfish farm. The shellfish were probably contaminated either in the submersible tanks or in a depuration land-based tank where they were stored. The source of contamination was not identified but shellfish could have been tainted by sewage overflows or by wastewater releases from a polluted storm sewer close to the shellfish farm or from on-site sanitation facilities. To prevent future hepatitis A outbreaks due to shellfish consumption from this area, hazards specific to each farm should be analysed. Timely information on sewage overflows should also be part of communities’ efforts regarding sewage collection and treatment.

PURPOSE OF THE STUDY:Hepatitis C virus genotyping is needed for treatment decision and monitoring. The results of a genotyping assay based on real-time PCR and TaqMan chemistry were compared with the results of NS5B region sequencing. MATERIALS AND METHODS:One hundred and two sera (genotypes 1-6) were tested. Amplification and detection of viral RNA were performed with the Abbott RealTime HCV Genotype II assay targeting 5'non-coded region (5'NC) for the identification of genotypes 1 to 6 and NS5B, for 1a and 1b subtypes detection. Sequencing of 5'NC fragment was used to resolve discrepant results. RESULTS:No indeterminate results were obtained. Concordance with NS5B sequencing was 93% (95 on 102), 96% at the genotype level (98 on 102) and 93% for genotype 1 subtyping (40 on 43). Discordant genotyping results were a 2f subtype identified as 5, a 6a typed as 1, a 3a identified as a 1-3 co-infection and a 4r identified as a 1-4 co-infection. Discordant subtyping results were 2 1b subtypes only typed as 1 and a 1e identified as 1a. CONCLUSION:Abbott RealTime HCV Genotype II assay is a rapid, automated and simple to interpret method for HCV genotyping. It allows the detection of possible mixed infections which might have a negative impact on therapeutic response. However, the discrepant results found in this small series underline the need for assay optimization.