Absence of the mecC gene in methicillin-resistant Staphylococcus aureus isolated from various clinical samples: The first multi-centered study in Turkey.

BACKGROUND: mecA is a predefined gene causing methicillin resistance in Staphylococcus aureus (S. aureus) isolates; however, it has been shown that some methicillin-resistant S. aureus (MRSA) strains do not carry this gene. Recently, in isolates found to be MRSA-positive but mecA-negative, a new resistance gene called mecC, which is a homolog of mecA, has been reported. This study aimed to investigate the mecC and mecA genes in MRSA strains isolated from different geographic regions in Turkey. METHODS: The sample of the study consisted of 494 MRSA strains isolated from seven geographical regions in Turkey between 2013 and 2016. The strains were obtained from 17 centers, comprising 13 university hospitals, three education and research hospitals, and one state hospital. Methicillin resistance in S. aureus strains was determined using the agar disk diffusion method with a cefoxitin disk and the agar dilution method with oxacillin. The mecC and mecA genes in MRSA strains was investigated by Polymerase Chain Reaction (PCR). RESULTS: Of the MRSA strains investigated, 47.9% were isolated from intensive care units. Concerning sample type, 36.7% were detected in the respiratory tract (tracheal aspirate, sputum, etc.), 24.8% in blood, 18.7% in skin and soft tissues, 9.3% in nasal swabs, 5.4% in urine, 4.1% in ears, and 1% in sterile body fluid. Using PCR, mecC was not identified in any of the S. aureus strains isolated from different clinical microbiology laboratories. mecA gene positivity was found in 315 of the MRSA strains (63.8%). Staphylococcal Cassette Chromosome mec (SCCmec) type was identified in 232 strains (46.9%), of which 136 (58.7%) were type II, 75 (32.4%) were type IV, 12 (5.1%) were type IIIb, six (2.5%) were type I, and three (1.3%) were type III. CONCLUSION: This is the first multi-centered study to investigate MRSA strains isolated from different regions in Turkey. The mecC gene was not detected in any of the MRSA strains. We believe that this study will constitute an important basis for monitoring possible future changes.

Three cases of Chronic Suppurative Otitis Media (CSOM) caused by Kerstersia gyiorum and a review of the literature.

New identification techniques such as gene sequencing and mass spectrometry have increased the incidence of novel agents such as Kerstersia gyiorum. As a new member of the Alcaligenaceae family, K. gyiorum was isolated from wounds, respiratory tract, urine specimens and most frequently from chronic suppurative otitis media (CSOM). We isolated three K. gyiorum strains from three CSOM cases over a one-year period. The strains were analyzed by mass spectrometry and identified by Bruker Biotyper 3.1 (Bruker Daltonics, USA). The cases were young patients without chronic diseases and immunodeficiencies. Two strains were resistant to ciprofloxacin.

In vitro susceptibility to methicillin, vancomycin and linezolid of staphylococci isolated from bloodstream infections in eastern Turkey.

Staphylococcus species are one of the major causes of bacterial bloodstream infections. Multi-resistant staphylococci infections are major therapeutic problems. This study was aimed to detect methicillin, linezolid and vancomycin susceptibilities of Staphylococcus isolates. A total of 870 Staphylococcus strains isolated from blood cultures of hospitalized patients with BSI. Antimicrobial susceptibilities of methicillin, linezolid and vancomycin were detected according to the Clinical and Laboratory Standards Institute (CLSI). A total of 771 (88.6%) isolates were coagulase-negative staphylococci (CoNS). 700 (80.5%) isolates were methicillin-resistant (MR) and 170 (19.5%) were methicillin-susceptible (MS). All the MS isolates were also susceptible to linezolid. However 15 (1.7%) of MR strains were resistant to linezolid. The minimum inhibitory concentration range for the linezolid-resistant isolates by Etest was 6-32 µg/mL. The difference between linezolid susceptibilities for MS and MR staphylococci was not quite statistically significant (p = 0.052). There was no statistically significant difference between S. aureus and CoNS isolates for linezolid susceptibility. All of the isolates were susceptible to vancomycin. In conclusion, linezolid is currently an efficient option for the treatment of methicillin-resistant staphylococci infections.

In vitro susceptibility to methicillin, vancomycin and linezolid of staphylococci isolated from bloodstream infections in eastern Turkey

Staphylococcus species are one of the major causes of bacterial bloodstream infections. Multi-resistant staphylococci infections are major therapeutic problems. This study was aimed to detect methicillin, linezolid and vancomycin susceptibilities of Staphylococcus isolates. A total of 870 Staphylococcus strains isolated from blood cultures of hospitalized patients with BSI. Antimicrobial susceptibilities of methicillin, linezolid and vancomycin were detected according to the Clinical and Laboratory Standards Institute (CLSI). A total of 771 (88.6%) isolates were coagulase-negative staphylococci (CoNS). 700 (80.5%) isolates were methicillin-resistant (MR) and 170 (19.5%) were methicillin-susceptible (MS). All the MS isolates were also susceptible to linezolid. However 15 (1.7%) of MR strains were resistant to linezolid. The minimum inhibitory concentration range for the linezolid-resistant isolates by Etest was 6-32 µg/mL. The difference between linezolid susceptibilities for MS and MR staphylococci was not quite statistically significant (p = 0.052). There was no statistically significant difference between S. aureus and CoNS isolates for linezolid susceptibility. All of the isolates were susceptible to vancomycin. In conclusion, linezolid is currently an efficient option for the treatment of methicillin-resistant staphylococci infections.

Effect of routine Hepatitis B vaccination program in Southeast of Turkey? Comparing of the results of HBV DNA in terms of age groups for the years 2002 and 2012.

Diyarbakir is the largest residential area in the Southeastern Anatolia of Turkey. Routine HBV vaccination has begun to be implemented by the Ministry of Health in Turkey in 1998. The purposes of this study were to detect the levels of HBV DNA in patients with HBV in 2012, and to compare the results of the year 2002 according to age groups. HBV DNA results of patients were divided in to seven age groups (0-14, 15-20, 21-30, 31-40, 41-50, 51-60, and > 61 years) and for comparison of HBV DNA levels of 2002 and 2012, HBV DNA values in pg/ml of year 2002 were translated into IU/ml and HBV DNA levels were grouped as < 5 pg/ml < 2.43 × 10(5) IU/ml, 5-100 pg/ml 2.43 × 10(5)-4.86 × 106 IU/ ml, 101-2000 pg/ml 4.87 × 10(6)-9.72 × 10(7) IU/ml, > 2000 pg/ml > 9.72 × 10(7) IU/ml 2-3. A statistically significant decrease was seen in the number of individuals in 0-14 age group in 2012 compared with 2002. In 2002 the rate of individuals in 0-14 age group was 18.8% whereas 4.8% in 2012. Our study was suggested that that routine HBV vaccination program, contributed to the reduced risk of HBV infection in our region.

Assessment of methicillin and clindamycin resistance patterns in Staphylococcus aureus isolated from a tertiary hospital in Turkey.

In order to detect the methicillin susceptibility and determine the prevalence of constitutive and inducible clindamycin resistance in Staphylococcus aureus strains, a total of 254 S. aureus isolates, including 139 (54.7%) methicillin-sensitive and 115 (45.3%) methicillin-resistant, were tested for clindamycin and erythromycin by Kirby-Bauer disk diffusion method. The disk diffusion induction test or D-test was performed on erythromycin-resistant and clindamycin-sensitive strains using the disk diffusion method. The erythromycin-resistant and clindamycin-sensitive strains with a D-shaped zone around the clindamycin disk were considered positive for inducible clindamycin resistance. If a strain was found to be both erythromycin-resistant and clindamycin-resistant, it was considered to have constitutive clindamycin resistance. Overall, ten (3.9%) strains, including two methicillin-resistant S. aureus and eight methicillin-sensitive S. aureus, were found to be erythromycin-resistant and clindamycin-sensitive according to the disk diffusion method and all of these strains showed inducible clindamycin resistance by the D-test method. Constitutive clindamycin resistance was detected in 100 of all MRSA strains. In conclusion, the presence of inducible clindamycin-resistant strains may lead to clindamycin treatment failure in patients with S. aureus infection. The D-test method should be used for antimicrobial susceptibility testing of clindamycin.

[Evaluation of rapid genotype assay for the identification of gram-positive cocci from blood cultures and detection of mecA and van genes]. / Kan Kültürlerinde Üreyen Gram-Pozitif Koklarin Tanimlanmasi ve mecA ve van Genlerinin Saptanmasinda Hizli Genotip Testinin Degerlendirilmesi.

Rapid and accurate identification of bacterial pathogens grown in blood cultures of patients with sepsis is crucial for prompt initiation of appropriate therapy in order to decrease related morbidity and mortality rates. Although current automated blood culture systems led to a significant improvement in bacterial detection time, more rapid identification systems are still needed to optimise the establishment of treatment. Novel genotype technology which is developed for the rapid diagnosis of sepsis, is a molecular genetic assay based on DNA multiplex amplification with biotinylated primers followed by hybridization to membrane bound probes. The aim of this study was to evaluate the performance of "Genotype® BC gram-positive” test for the identification of gram-positive cocci grown in blood cultures and rapid detection of mecA and van genes. This test uses DNA.STRIP® technology which includes a panel of probes for identification of 17 gram-positive bacterial species and is able to determinate the methicillin and vancomycin resistance mediating genes (mecA and vanA, vanB, vanC1, vanC2/C3) simultaneously, in a single test run. A total of 55 positive blood cultures from BACTECTM Plus/F (Becton Dickinson, USA) aerobic and pediatric blood culture vials were included in the study. The isolates which exhibit gram-positive coccus morphology by Gram staining were identified by Genotype ® BC gram-positive test (Hain Life Science, Germany). All of the samples were also identified with the use of Phoenix PMIC/ID Panel (Becton Dickinson, USA) and antibiotic susceptibilities were determined. Of the 55 blood culture isolates, 17 were identified as Staphylococcus epidermidis [all were methicillin-resistant (MR)], 9 were S.aureus (one was MR), 18 were S.hominis (10 were MR), 4 were E.faecalis, 3 were E. faecium (one was vanconycin-resistant), 2 were S.saprophyticus (one was MR), 1 was S.warneri and 1 was S.haemolyticus, by Phoenix automated system. Genotype® BC gram-positive test results revealed consistency with Phoenix system regarding bacterial identification in 46 (83.6%) of the samples. The two bacteria identified as S.saprophyticus by the Phoenix system could not be identified by the Genotype® BC test since this species were not included in the identification panel of the system, however, mecA gene were detected in these two samples by Genotype® BC test. Genotype® BC test detected mecA gene in five samples which were not detected as methicillin resistant by the Phoenix system. Besides polymicrobial growth was determined in five samples by Genotype ® BC test, but not by the automated system. One E.faecium isolate with vanA gene was correctly identified by Genotype® BC test. In conclusion, Genotype® BC gram-positive test is a fast and reliable test for the identification of the most important gram-positive pathogens and mecA and van genes directly from positive blood culture bottles. This test was also found superior than the automated Phoenix system regarding the detection of polymicrobial growth. These data indicated that, routine use of DNA strip technology-based assay would be useful for clinical diagnosis in patients with sepsis.

[Investigation of anticardiolipin antibodies in chronic hepatitis B infection together with total anti-delta positivity]. / Kronik hepatit B enfeksiyonu ile birlikte total anti-delta pozitifliginde antikardiyolipin antikorlarinin arastirilmasi.

Anticardiolipin antibodies (ACAs) are formed against phospholipids in various clinical conditions such as autoimmune diseases, malignancy, infectious diseases, alcohol-related and hepatic cirrhosis. The aims of this study were to investigate the presence of ACAs in patients with chronic hepatitis B together with positive total anti-delta antibodies, and to investigate the relationship between age, gender, and some laboratory parameters (ALT, AST, albumin, globulin, platelet number) of patients with chronic hepatitis delta virus (HDV) infection, who were positive or negative for ACAs. A total of 60 patients (43 male, 17 female) with chronic hepatitis D infection [HBsAg positive, HBeAg negative, anti-HBe positive, anti-HBc IgG positive, anti-HBc IgM negative, total anti-delta positive, anti-HCV negative] and 30 patients (21 male, 9 female) without hepatitis D infection [HBsAg positive, HBeAg negative, anti-HBe positive, anti-HBc IgG positive, anti-HBc IgM negative, total anti-delta negative, anti-HCV negative] as control group were included to the study. ACA IgG and IgM were searched by a commercial microELISA kit (Euroimmun, Germany). The statistical evaluation was performed with Pearson's chi-square test, Student's t-test, and Fisher's exact test. Total ACAs positivity rate of 60 patients with chronic HDV infection, was found as 13.3%, in which four of the patients were positive for only ACA IgM, while four was positive for only IgG. Positivity for both ACA IgG and ACA IgM could not be detected in these patients. No patients in the control group had positivity for ACAs (IgG and/or IgM). A statistically significant difference was observed in terms of ACA positivity between patients with and without HDV infection (p< 0.05). After all, there was no statistically significant correlation between ACAs positivity and the age, sex, and laboratory parameters of the patients with chronic HDV infection, except lower serum albumin levels (p= 0.004). Although the data of this study revealed a statistically significant positive correlation between chronic HDV infection and anticardiolipin antibodies, it is clear that there is a need for further studies on this subject.

Oxybutynin effects on Staphylococcus epidermidis biofilm production.

BACKGROUND: Oxybutynin is a tertiary amine and has chemical similarities like protamine sulfate. Protamine sulfate's effect on bacterial viability has been shown in some studies; however, there is scanty data regarding the effect of oxybutynin on bacterial viability. Accordingly, the aim of the present study was to investigate the effect of oxybutynin on bacterial viability. METHODS: Twenty of Staphylococcus epidermidis (S. epidermidis) strains were isolated from patients who had catheter-associated urinary tract infection. These strains were exposed to 5 different concentrations of oxybutynin in tryptic soy broth (TSB) media (max: 5 mg/mL, min: 0.05 mg/mL). Quantitative growth patterns were measured by spectrophotometer. Under the same media conditions, biofilm production of individual strains was measured by Deighton's micromethod at the end of the 6-hour incubation period. RESULTS: After the incubation period, weak and strong slime-producing bacteria groups were obtained and both groups' slime productions were extensively low at 5 mg/mL concentration of oxybutynin. Slime production of S. epidermidis was inversely correlated with oxybutynin concentrations. CONCLUSION: Although there was a dose-dependent reduction of biofilm production, there seemed to be no bactericidal effect of oxybutynin on S. epidermidis.

[Investigation of autoantibody, anti-HCV and anti-HIV seropositivities in "anti-HBc alone" positive samples]. / SALT Anti-HBc Pozitif orneklerde otoantikor, anti-HCV ve anti-HIV seropozitifliginin arastirilmasi.

"Anti-HBc alone" which is an unusual serologic pattern of hepatitis B virus (HBV) infections, may be detected in the seropositive samples for hepatitis C virus (HCV), human immunodeficiency virus (HIV) infections and in the presence of autoantibodies due to cross reactions. In this study, 20 serum samples with isolated antibody to hepatitis B core antigen, which were detected in May 2005, have been investigated by means of the presence of some autoantibodies (anti-nuclear antibody; ANA and rheumatoid factor; RF), anti-HCV and anti-HIV, in the Central Laboratory of Dicle University Medical School. All of the "anti-HBc alone" samples were negative for HBV-DNA by real-time polymerase chain reaction (PCR), and liver enzyme (ALT and AST) levels were normal except for three patients. As a result, a total of six (30%) samples were found positive. Four of them were positive for ANA and two were positive for anti-HCV, while one serum yielded positivity for both ANA and anti-HCV. Anti-HCV positive samples were searched for the presence of HCV-RNA by real-time PCR, and none were found positive. Of three patients with increased AST and ALT levels, one was anti-HCV positive, one was ANA positive, while the other was negative for all parameters. In conclusion, possible presence of autoantibodies and anti-HCV should be taken into consideration during the evaluation of isolated anti-HBc IgG positive test results.

[Short communication: evaluation of the correlation between hepatitis D virus (HDV) RNA positivity and HDV antibodies]. / Kisa bildiri: hepatit D virus (HDV) RNA pozitifligi ile HDV antikorlari arasindaki iliskinin degerlendirilmesi.

The objective of this study was to evaluate the correlation between serum hepatitis D -delta- virus (HDV) RNA detection and anti-HDV IgG and IgM antibodies, in the serodiagnosis of delta hepatitis. A total of 153 HBsAg positive sera were screened for the presence of anti-HBc IgM, anti-HDV IgG and anti-HDV IgM by commercial enzyme immunoassays and HDV-RNA by real time polymerase chain reaction (PCR). Of 153 sera, 86 (56.2%) were found positive for HDV antibodies. Although isolated anti-HDV IgG was present in 35 and isolated anti-HDV IgM was present in 11 patients, IgG and IgM were present concurrently in 40 additional patients. HDV-RNA was detected in 21.5% (33/153) of the patients. Four of the 33 HDV-RNA positive patients were positive only for anti-HDV IgG, 8 were positive only for anti-HDV IgM, and 19 were positive for both anti-HDV IgG and IgM antibodies. Twenty seven of 51 (53%) anti-HDV IgM positive patients were also found positive for HDV-RNA, while 27 of 33 (82%) HDV-RNA positive patients exhibited anti-HDV IgM positivity. Increased serum ALT levels were detected approximately in 85% (28/33) of viremic patients. As all of the HDV-RNA positive patients were found negative for anti-HBc IgM, superinfection with delta virus were considered. In conclusion, PCR is a sensitive and useful method for the detection of viremic patients as well as for the monitorization of antiviral therapy, anti-HDV IgM positivity together with increased ALT levels appear to be good markers for the prediction of hepatitis delta viremia, especially in the countries with limited economical sources as Turkey.

[Comparison of microscopy and EIA in the diagnosis of Giardia intestinalis in stool specimens.]. / Giardiasisin tanisinda Enzym Immun Assay (EIA) ve direkt inceleme yöntemlerinin karsilastirilmasi

In this study; we compared the direct microscopic method and EIA test in the investigation of the stools of patients with gastrointestinal symptoms who presented at clinics. A total of 188 stool specimens collected from clinics were investigated by direct microscopy using native-Lugol preparations. Giardia cysts and/or trophozoites were observed in 141 specimens. There were no Giardia cysts and/or trophozoites or any other intestinal parasites detected in the other 47 stool specimens. The RIDASCREENR EIA kit procedure was applied in all specimens. Out of 141 specimens positive with direct microscopy, 136 specimens were positive with the EIA test and 5 specimens, negative. Parasites were not found in 47 stool specimens with direct microscopy. Of these, 38 specimens were negative with the EIA test and 9 specimens, positive. When the patient and control groups were compared, a significant difference was observed between the two methods (p < 0.05). The sensitivity and specificity of the EIA method that was used to determine the antigenic properties of G. intestinalis in stools were 96.4% and 80.8%, respectively.

The seroprevalence of varicella zoster antibodies in Behçet's and other skin diseases.

The purpose of this study was to investigate the association of the varicella zoster virus (VZV) IgG, and IgM antibodies with Behçet's and other skin diseases (group 1: recurrent aphthos stomatitis, fungal infections, psoriasis; group 2: vitiligo, lichen planus). Twenty eight patients with Behçet's disease (BD), and 117 patients with dermatological disorders other than BD were evaluated for specific VZV IgG and IgM antibodies by using a third generation enzyme-linked immunosorbent assay (ELISA). The Mantel Heizshel chi2 method was used to adjust the confounding of age and sex of the patients. The serological positivity for VZV IgG and IgM antibodies in BD was not statistically different from other skin diseases. When we considered the age of the patients, chi2 = 2.64, CI (0.27-1.65), odds ratio (1, 1.25, 2.21) (p = 0.10) and when we considered the sex of the patients, chi2 = 0.31, CI (0.81-1.28), odds ratio (1, 1.45, 1.41), (p = 0.57).