Spread of imipenem-resistant Acinetobacter baumannii co-expressing OXA-23 and GES-11 carbapenemases in Lebanon.

OBJECTIVES: The acquisition of carbapenemases by Acinetobacter baumannii is reported increasingly worldwide, but data from Lebanon are limited. The aims of this study were to evaluate the prevalence of imipenem-resistant A. baumannii in Lebanon, identify resistance determinants, and detect clonal relatedness. METHODS: Imipenem-resistant A. baumannii were collected from nine Lebanese hospitals during 2012. Antimicrobial susceptibility, the cloxacillin effect, and ethylenediaminetetraacetic acid (EDTA) synergy were determined. Genes encoding carbapenemases and insertion sequence ISAba1 were screened via PCR sequencing. ISAba1 position relative to genes encoding Acinetobacter-derived cephalosporinases (ADCs) and OXA-23 was studied by PCR mapping. Clonal linkage was examined by enterobacterial repetitive intergenic consensus PCR (ERIC-PCR). RESULTS: Out of 724 A. baumannii isolated in 2012, 638 (88%) were imipenem-resistant. Of these, 142 were analyzed. Clavulanic acid-imipenem synergy suggested carbapenem-hydrolyzing extended-spectrum ß-lactamase. A positive cloxacillin test indicated ADCs, while EDTA detection strips were negative. Genotyping indicated that 90% of isolates co-harbored blaOXA-23 and blaGES-11. The remaining strains had blaOXA-23, blaOXA-24, blaGES-11, or blaOXA-24 with blaGES-11. ISAba1 was located upstream of blaADC and blaOXA-23 in 97% and 100% of isolates, respectively. ERIC-PCR fingerprinting revealed 18 pulsotypes spread via horizontal gene transfer and clonal dissemination. CONCLUSION: This survey established baseline evidence of OXA-23 and GES-11-producing A. baumannii in Lebanon, indicating the need for further surveillance.

Countrywide spread of OXA-48 carbapenemase in Lebanon: surveillance and genetic characterization of carbapenem-non-susceptible Enterobacteriaceae in 10 hospitals over a one-year period.

OBJECTIVES: To detect, characterize, and assess the genetic clonality of carbapenem-non-susceptible Enterobacteriaceae in 10 Lebanese hospitals in 2012. METHODS: Selected Enterobacteriaceae isolates with reduced susceptibility to carbapenems were subject to phenotypic study including antibiotic susceptibility, cloxacillin effect, modified Hodge test, and activity of efflux pump inhibitor. Carbapenemase genes were detected using PCR; clonal relatedness was studied by pulsed field gel electrophoresis. RESULTS: Out of 8717 Enterobacteriaceae isolated in 2012, 102 (1.2%) showed reduced susceptibility to carbapenems. Thirty-one (70%) of the 44 studied clinical isolates harbored blaOXA-48, including 15 Klebsiella pneumoniae, eight Escherichia coli, four Serratia marcescens, three Enterobacter cloacae, and one Morganella morganii. The majority of OXA-48 producers co-secreted an extended-spectrum beta-lactamase, while one had an acquired AmpC of the ACC type. In the non-OXA-48 producers, carbapenem resistance was attributed to the production of acquired AmpC cephalosporinases of MOX or CIT type, outer membrane impermeability, and/or efflux pump overproduction. DNA fingerprints revealed that OXA-48 producers were different, except for clonal relatedness among four K. pneumoniae, two E. coli, two E. cloacae, and three S. marcescens. CONCLUSIONS: Nosocomial carbapenem-non-susceptible Enterobacteriaceae are moderately spread in Lebanon and the predominant mechanism is OXA-48 production.

Contribution of common variants of ENPP1, IGF2BP2, KCNJ11, MLXIPL, PPARγ, SLC30A8 and TCF7L2 to the risk of type 2 diabetes in Lebanese and Tunisian Arabs.

BACKGROUND: While several type 2 diabetes mellitus (T2DM) susceptibility loci identified through genome-wide association studies (GWAS) have been replicated in many populations, their association in Arabs has not been reported. For this reason, the present study looked at the contribution of ENNP1 (rs1044498), IGF2BP2 (rs1470579), KCNJ11 (rs5219), MLXIPL (rs7800944), PPARγ (rs1801282), SLC30A8 (rs13266634) and TCF7L2 (rs7903146) SNPs to the risk of T2DM in Lebanese and Tunisian Arabs. METHODS: Study subjects (case/controls) were Lebanese (751/918) and Tunisians (1470/838). Genotyping was carried out by the allelic discrimination method. RESULTS: In Lebanese and Tunisians, neither ENNP1 nor MLXIPL was associated with T2DM, whereas TCF7L2 was significantly associated with an increased risk of T2DM in both the Lebanese [P < 0.001; OR (95% CI): 1.38 (1.20-1.59)] and Tunisians [P < 0.001; OR (95% CI): 1.36 (1.18-1.56)]. Differential associations of IGF2BP2, KCNJ11, PPARγ and SLC30A8 with T2DM were noted in the two populations. IGF2BP2 [P = 1.3 × 10(-5); OR (95% CI): 1.66 (1.42-1.94)] and PPARγ [P = 0.005; OR (95% CI): 1.41 (1.10-1.80)] were associated with T2DM in the Lebanese, but not Tunisians, while KCNJ11 [P = 8.0 × 10(-4); OR (95% CI): 1.27 (1.09-1.47)] and SLC30A8 [P = 1.6 × 10(-5); OR (95% CI): 1.37 (1.15-1.62)] were associated with T2DM in the Tunisians, but not Lebanese, after adjusting for gender and body mass index. CONCLUSION: T2DM susceptibility loci SNPs identified through GWAS showed differential associations with T2DM in two Arab populations, thus further confirming the ethnic contributions of these variants to T2DM susceptibility.

Is thrombophilia a major risk factor for deep vein thrombosis of the lower extremities among Lebanese patients?

AIM: Factor V Leiden (R506Q) mutation is the most commonly observed inherited genetic abnormality related to vein thrombosis. Lebanon has one of the highest frequencies of this mutation in the world with a prevalence of 14.4% in the general population. The aim of this study is to define risk factors including inherited genetic abnormalities among Lebanese patients with lower extremity deep vein thrombosis. We report the clinical outcome of patients with thrombophilia. METHODS: From January 1998 to January 2008, 162 patients (61 males and 101 females) were diagnosed with lower extremity deep vein thrombosis. Mean age was 61 years (range: 21 to 95 years). RESULTS: The most frequent risk factors for vein thrombosis were surgery, advanced age, obesity, and cancer. Twenty-five patients had thrombophilia, 16 patients had factor V Leiden (R506Q) mutation, and seven patients had MTHFR C677T mutation. Ninety-two percent of patients screened for thrombophilia were positive. Screening was requested in young patients (16), patients with recurrent (11), spontaneous (8), and extensive (5) venous thrombosis, familial history (5), pregnancy (4), estroprogestative treatment (3), and air travel (1). Nine patients had one, 11 patients had two, and five had three of these conditions. Follow-up (6 to 120 months) of these 25 patients treated with antivitamin K did not reveal recurrences or complications related to venous thromboembolism. CONCLUSION: Factor V Leiden mutation followed by MTHFR mutation are the most commonly observed genetic abnormalities in these series. Defining risk factors and screening for thrombophilia when indicated reduce recurrence rate and complications. Recommendations for thrombophilia screening will be proposed.

Antimicrobial susceptibility patterns of Haemophilus influenzae and Streptococcus pneumoniae isolates in a Beirut general university hospital between 2000 and 2004.

Susceptibility patterns of Streptococcus pneumoniae and Haemophilus influenzae collected over a 5-year period in a Beirut general university hospital were studied. Only 40.6-50% of S. pneumoniae isolates were susceptible to penicillin G. Susceptibility to clindamycin and erythromycin decreased from 94.1% and 89.7%, respectively, in 2000 to 75% and 71.9%, respectively, in 2004. All isolates were susceptible to ceftriaxone, ciprofloxacin and vancomycin. For H. influenzae, no resistance was observed to amoxycillin-clavulanate, ceftriaxone, ciprofloxacin and rifampicin, with >92% of isolates showing susceptibility to cefuroxime, chloramphenicol, erythromycin and tetracycline. The proportion of beta-lactamase-positive isolates varied between 22.7 and 30.8%.

[Prevalence and susceptibility patterns of extended-spectrum betalactamase-producing Escherichia coli and Klebsiella pneumoniae in a general university hospital in Beirut, Lebanon]. / Prevalencia y patrones de sensibilidad de Escherichia coli y Klebsiella pneumoniae productoras de betalactamasas de espectro ampliado en Beirut, Líbano.

Extended-spectrum betalactamases (ESBLs) are recognized worldwide as a problem in hospitalized patients. Their prevalence among clinical isolates of Enterobacteriaceae varies between countries and institutions. We studied the evolution of ESBL production by clinical isolates of Escherichia coli and Klebsiella pneumoniae and analyzed the patterns of susceptibility of these isolates to different antimicrobial agents in a general university hospital in Beirut. Of the 4299 isolates of E. coli and 1248 isolates of K. pneumoniae tested over the five years, 2.0% of the E. coli and 20.0% of K. pneumoniae were ESBL producing. A clear decrease in the susceptibility to all antibiotics was observed between 1999 and 2001, and no resistance to imipenem was detected. The isolates were distributed between the Intensive Care Unit (ICU), medical wards, outpatients, and other origins. The highest numbers were found in the ICU (E. coli 28.1% and K. pneumoniae 34.8%). Three phenotypes of resistance to cefotaxime and ceftazidime were observed on the basis of microbiological results. The present study was the first to assess the occurrence and susceptibility patterns of extended-spectrum betalactamase-producing Enterobacteriaceae in Lebanon.

Prevalencia y patrones de sensibilidad de Escherichia coli y Klebsiella pneumoniae productoras de betalactamasas de espectro ampliado en Beirut, Líbano / Prevalence and susceptibility patterns of extended-spectrum betalactamase-producing Escherichia coli and Klebsiella pneumoniae in a general university hospital in Beirut, Lebanon

Las betalactamasas de espectro ampliado (BLEA) representan un problema de ámbito mundial en los pacientes hospitalizados. Su prevalencia entre las cepas clínicas de Enterobacteriaceae varía según los países y las instituciones. Estudiamos la evolución de la producción de BLEA por cepas clínicas de Escherichia coli y Klebsiella pneumoniae y analizamos sus patrones de sensibilidad a distintos agentes antimicrobianos en un hospital general universitario en Beirut. De las 4299 cepas de E. coli y las 1248 de K. pneumoniae analizadas durante cinco años, un 2 por ciento de las cepas de E. coli y un 20 por ciento de las de K. pneumoniae fueron productoras de BLEA. Entre 1999 y 2001 se observó una clara disminución de la sensibilidad frente a todos los antibióticos y no se detectó ninguna resistencia al imipenem. Las cepas fueron aisladas en la Unidad de Cuidados Intensivos (UCI), las salas hospitalarias, las consultas externas y otros lugares. La mayor cantidad de cepas se aisló en la UCI (un 28,1 por ciento de E. coli y un 34,8 por ciento de K. pneumoniae). En el estudio microbiológico se identificaron tres fenotipos de resistencia a cefotaxima y ceftazidima. El presente estudio es el primero en evaluar la existencia y los patrones de sensibilidad de enterobacterias productoras de BLEA en Líbano (AU)

Prevalence of antibodies against hepatitis C virus among blood donors in Lebanon, 1997-2000.

Chronic hepatitis C virus (HCV) infection in many individuals is asymptomatic and the prevalence of antibodies to hepatitis C virus (anti-HCV) among blood donors in Lebanon is scarce. This study aimed to address the prevalence of anti-HCV in 8700 blood donors, the data obtained was compared to other world regions. Between 1997 and 2000, 8700 blood donors were screened for the presence of anti-HCV in their sera. Initially reactive specimens were retested in duplicate, and repeatedly positive samples were subsequently retested by a third generation microplate enzyme immunoassay. Of the 8700 blood donors screened, 51 were confirmed positive for anti-HCV, giving a prevalence rate of 0.6%. While there was no difference in anti-HCV prevalence in relation to age or gender, higher rates were seen in non-Lebanese compared to Lebanese subjects (6.17% vs. 0.48%, P < 0.001). None of the anti-HCV positive individuals had an identifiable risk factor for contracting HCV (intravenous drug user, prior transfusion, etc.), and their transaminases were comparable to anti-HCV-negative donors, suggesting that HCV-positive donors were asymptomatic. These results demonstrate low prevalence of anti-HCV among Lebanese blood donors, which was comparable to those established for Western countries.

[Post-varicella thrombosis and factor V Leiden mutation]. / Thrombose post-varicelle et mutation du facteur V Leiden.

BACKGROUND: Varicella is a benign infectious disease that is rarely complicated by thrombosis. Factor V Leiden (FVL) mutation is presently recognized as the most common inherited cause of thrombosis. From a case report the link between varicella associated thrombosis and FVL is discussed. CASE REPORT: An eight year-old boy was admitted on 15th day of a varicella infection for difficulty to walk and painful legs. Doppler ultra sound of the lower limbs venous system showed thrombosis of the left iliac vein and femoral veins. Thrombophilia work-up revealed a FVL heterozygotic status for the patient. Biological screening for molecular abnormalities associated with thrombophilia led to the detection of a heterozygous inherited FVL mutation in the patient. Response to heparine followed by oral anticoagulant treatment was good. CONCLUSION: Thrombosis associated with varicella in child is a rare event. We recommend to realize a thrombophilia screening in such patients. We also support varicella vaccination of children identified as having constitutional prothrombotic disorders like FVL mutation.

Molecular diagnosis of human papillomavirus: comparison between cervical and vaginal sampling.

BACKGROUND: Human papillomavirus (HPV) is the most significant cause of cervical cancer. In view of the number of drawbacks associated with endocervical sampling, the gold standard for HPV detection, this study examined the utility and specificity of vaginal sampling as an alternative for endocervical sampling for the routine detection of HPV. CASE STUDY: The study comprised 51 women who tested positive and 54 women who tested negative for endocervical HPV by polymerase chain reaction (PCR), confirmed by histopathology. At the time of specimen collection, both (speculum-assisted) endocervical and vaginal (no speculum) scrapings were isolated from HPV-positive and negative women, and HPV DNA was assessed by PCR using the MY09/MY11 primer system; HPV type was identified by hybridization of PCR products with type-specific biotinylated DNA probes. Each participant served as her own control. HPV was detected in vaginal and cervical scrapes from all HPV-positive but not HPV-negative women. In HPV-positive women the same HPV type was found in vaginal and endocervical scrapings (positive predictive value = 1.0). CONCLUSION: Correlation between vaginal and endocervical sampling methods was excellent in detecting the presence of HPV DNA and for identifying distinct HPV genotypes. Utilization of vaginal testing for routine HPV detection, and for the long-term follow-up of persistent HPV infection, is therefore recommended.

Seroprevalence of hepatitis C virus (HCV) infection among blood donors: a hospital-based study.

Insofar as chronic hepatitis C virus (HCV) infection in many individuals is asymptomatic, and as the prevalence of antibodies to hepatitis C virus (anti-HCV) among blood donors in Lebanon is scarce, this study addressed the prevalence of anti-HCV in 5,115 blood donors. Data obtained were compared to other world regions. Of the blood donors screened, 57 were initially tested positive or doubtful for anti-HCV Ab. Subsequent testing by two-third generation enzyme immunoassays confirmed that, of the 57 initially tested positive/doubtful, only 18 were positive for anti-HCV giving a prevalence rate of 0.4%. While there was no difference in HCV prevalence with respect to age or gender, a higher rate was seen in non-Lebanese compared to Lebanese subjects (3.4% vs 0.3%, P < 0.001). These results demonstrate a low prevalence of HCV infection among Lebanese blood donors, which was comparable to those established for western countries.

Factor V R506Q mutation-Leiden: an independent risk factor for venous thrombosis but not coronary artery disease.

BACKGROUND: A specific point G-A transition at nucleotide position 1691 in the factor V (FV) gene, FV-Leiden, was associated with increased risk of venous thromboembolism (VTE). Insofar as the association of FV-Leiden with coronary artery disease (CAD) remains poorly defined, the aim of this study was to determine the prevalence of FV-Leiden in a sample of 68 VTE patients, 69 CAD patients, and 192 randomly selected healthy subjects. METHODS: Total genomic DNA was extracted from the peripheral blood of study subjects and was used for PCR analysis. The presence (or absence) of FV-Leiden was assessed by PCR using primers flanking the mutant site (nt 1691), followed by hybridization with wild-type ('G') and mutant ('A') biotinylated DNA probes; detection was by DNA enzyme immunoassay (DEIA). RESULTS: While the prevalence of FV-Leiden in CAD patients was not statistically different from that of healthy subjects (14.5 % vs. 15.1 %; P=0.890, odds ratio 0.95; 95 % confidence interval 0.43--2.06), a significant increase in FV-Leiden prevalence was seen in VTE patients (70.6 % in VTE patients; P<0.001, odds ratio 13.4, 95 % confidence interval 6.9--25.8). Of the 48 VTE patients who tested positive for FV-Leiden, 42 were heterozygotes (G/A), while 6 were homozygotes (A/A) (allele frequency 0.397). All 10 CAD patients positive for FV-Leiden were heterozygote carriers (allele frequency 0.072). While gender was not a factor in FV-Leiden expression, higher prevalence in FV-Leiden was seen in younger (< or =45 years) VTE patients (38/51 vs. 10/17). CONCLUSION: FV-Leiden is a major inherited risk factor for VTE, with a peak incidence in younger patients, but does not appear to play any role in CAD pathogenesis in the population studied.

Seroprevalence of hepatitis C infection among health care personnel in Beirut, Lebanon.

BACKGROUND: Health care workers are at risk of contracting hepatitis C virus (HCV) infection more than the general population, and chronic HCV infection may be asymptomatic. Therefore, the purpose of this study was to determine the seroprevalence of HCV among health care workers at St Georges-Orthodox Hospital, the first study done for a major teaching hospital in Lebanon. METHODS: Health care personnel at St Georges-Orthodox Hospital, Beirut, were offered anonymous testing for anti-HCV antibody. Seroprevalence rates of health care personnel were compared with the rates of blood donors screened during the same year. RESULTS: Of the 502 persons screened, 13 (2.60%) initially tested either positive or doubtful-positive by the SM-HCV rapid test; 2 (0.4%) were confirmed positive by 2 commercial enzyme-linked immunosorbent assay kits and reverse transcriptase-polymerase chain reaction. This prevalence rate was comparable with the rate obtained for blood donors (n = 600) during the same period. CONCLUSION: The seroprevalence of HCV infection among health care workers at St Georges-Orthodox Hospital was similar to the rate observed in local blood donors, which suggests that the occupational risk of HCV infection was low.

Detection of human papillomavirus (HPV) genotypes in cervico-vaginal scrapes of women with normal and abnormal cytology.

BACKGROUND: Certain types of human papillomavirus (HPV) are associated with cervical intraepithelial neoplasia (CIN) and invasive cervical carcinoma. The study addressed the expression and detection of HPV genotypes in cervical and vaginal specimens of women with normal and abnormal cytology by polymerase chain reaction (PCR), each woman serving as her own control. METHODS: Study participants (127) were subgrouped into CIN-positive and CIN-negative, based on cytology screening, and endocervical and vaginal scrapes were collected by a gynecologist and placed immediately in saline. HPV DNA was assessed by PCR, and HPV genotypes were determined by hybridization of PCR products with type-specific biotinylated probes. RESULTS: Of the 127 participants, 55 tested positive and 72 tested negative for HPV DNA. While there was no difference between the two groups with regards to age or to number of pregnancies, higher numbers of smokers and of women with multiple sexual partners and abnormal cytology were seen in the HPV-positive group (P < 0.001). HPV DNA was detected in the vaginal scrapes of all HPV-positive, but in none of the HPV-negative women (sensitivity and specificity = 1.0). Furthermore, the HPV genotype was the same in vaginal and endocervical specimens in all the HPV-positive women. CONCLUSION: HPV detection by PCR, using endocervical or vaginal sampling, is a sensitive and highly specific test for the identification of HPV infection, in particular in women with cytomorphologically normal cervices.

Validity of vaginal testing in detecting human papillomavirus (HPV) genotypes.

The aim of this study was to assess the validity and usefulness of vaginal scrapes in detecting cervical human papillomavirus (HPV) DNA by the polymerase chain reaction (PCR). The study group comprised 23 women tested positive and 28 women tested negative for cervical HPV DNA by PCR, and confirmed by histopathology. At the time of specimen collection, both vaginal and endocervical scrapes were taken from these women, and tested for HPV DNA by PCR, using MY09/MY11 primer system. HPV genotypes were analyzed by hybridizing PCR products with HPV type-specific biotinylated probes. HPV DNA was detected in both vaginal and cervical scrapes from the HPV-positive, but not from HPV-negative group. In the HPV-positive group, the same HPV type was found in vaginal and endocervical scrapes, giving a positive predictive value of 1.0. The results indicate that HPV types can be detected in vaginal scrapes, and recommend utilization of the less invasive vaginal testing for the routine detection of HPV DNA.

The prevalence of factor V R506Q mutation-Leiden among apparently healthy Lebanese.

Resistance to activated protein C (APC) degradation caused by a specific point mutation in the factor V (FV) gene, FV:R506Q or FV-Leiden, which replaces Arg506 with Gln at the APC cleavage site within the FV gene, is the most prevalent inherited caused of venous thrombosis. Recent reports suggested that the prevalence of FV-Leiden is high among Caucasians, and very low among non-Caucasians, thereby suggesting that FV-Leiden has originated as a single event in a primary focus. Insofar as FV-Leiden is associated with increased risk of thromboembolic diseases, coupled with its selective worldwide distribution, the aim of this study was to determine the prevalence of FV-Leiden in Lebanon and compare it with those of other countries of Caucasian and non-Caucasian origin. FV-Leiden was determined by PCR, followed by hybridization with specific wild-type and mutant DNA probes. By screening 174 healthy Lebanese subjects, 25 were shown to carry the FV-Leiden mutation, giving an allele frequency of 7.4% and an overall prevalence rate of 14.4%. Of the 25 FV-Leiden carriers, 24 were in the heterozygous state while one was in the homozygous states. No statistical difference in the FV-Leiden prevalence was noted with respect to age, gender, or sect. In addition to Lebanon, which had the highest prevalence rate reported thus far (14.4%), a high prevalence of FV-Leiden was reported for Syria (13.6%), Greece-Cyprus (13.4%), and Jordan (12.3%), an indication that the Eastern Mediterranean is the primary focus of FV-Leiden mutation. The high prevalence of FV-Leiden in Lebanon suggests that screening for this mutation must be considered for those with a family history, and/or those with additional risk factors for venous thrombosis.