It is essential to evaluate the susceptibility of women in the reproductive age group to rubella virus in order to set strategies for the prevention of congenital rubella syndrome (CRS). Turkey began implementing measles-mumps-rubella vaccination as part of the national vaccination schedule for children (12 months, 6 years) and adolescents (14 years) in July, 2006, and there is an ongoing discussion of the need for a policy of vaccinating women of child-bearing age against rubella. The aim of this study was to determine the rubella seroprevalence among women in the reproductive age group in a rural district in Ankara and to provide data about rubella susceptibility for policymakers. Four hundred ninety of the women in the 15- to 49-year-old age group in the region who were targeted were reached (68.2%), and 467 (65.0%) of them who had a convenient serology were included in the study. Rubella IgG antibodies were quantified by the enzyme-linked immunosorbent assay method. Seropositivity was 95.5% for the total group and 96.2% among pregnant women. The seropositivity of this rural group of women was found to be high, but in order to rule out the need for a rubella vaccination program for women of child-bearing age, large-scale studies in different settings and studies that describe the CRS burden in Turkey are required.

The clinical efficacy of the monotherapy involving the administration of a high dose of dihydroartemisinin (DHA 900 mg) for 5 days was compared with that of the combination regimen (DHA 600 mg + mefloquine [MQ] 750 mg) in an open randomized study in 90 patients with uncomplicated falciparum malaria in the southern part of Viet Nam. Patients were randomly treated with the DHA-5 day monotherapy regimen (300, 300, 100, 100, and 100 mg given at 0, 24, 48, 72, and 96 h) or the DHA-MQ combination regimen (300 mg DHA at 0 h, then 300 mg DHA plus 750 mg MQ at 24 h). The end points for comparison were the parasite and fever clearance times (PCT and FCT) and recrudescence rates (by day 28 for DHA-5 days and day 42 for DHA-MQ). Eighty-nine patients completed the trial per protocol, including 45 cases receiving DHA-5 day and 44 receiving DHA-MQ. There was no difference in clinical manifestations, parasitemia density or other laboratory tests between the two patient groups. The PCTs were 35.3 ± 17.4 h (mean ± SD; range, 12 - 96) and 37.8 ± 19.2 h (range, 12 - 96), respectively for the DHA-5 day and DHA-MQ regimens (P > 0.05). Twelve patients receiving the DHA-5 day regimen relapsed with falciparum malaria by day 28 (26.7%) and 5 patients receiving the DHA-MQ regimen relapsed by day 42 (11.4%) (P = 0.07). Survival analysis showed that the DHA-5 day regimen had a radical cure rate significantly lower than that of the DHA-MQ regimen (P = 0.003). The high dose of DHA in the monotherapy regimen did not increase the efficacy of the treatment of patients with uncomplicated Plasmodium falciparum malaria. The DHA combination regimens are suggested to be the better regimens for DHA.

Intrauterine infection of the fetus is clearly an important mode of vertical transmission of hepatitis B virus (HBV). The trophoblast layer of the human placenta must be traversed by HBV in order to reach underlying cells and fetal capillaries. Although HBV has been detected in the trophoblast layer in situ, the degree of susceptibility of primary trophoblast cells to direct HBV infection in vitro remains unknown. To determine the receptivity of trophoblast cells to HBV infection and to discover the cellular basis for the molecular mechanism responsible for the passage of HBV from the maternal to the fetal circulation, we infected choriocarcinoma JEGIII cells and primary trophoblast cells with HBV. Our findings suggest that the cells could be reproducibly infected with HBV and that the infective patterns of the isolated trophoblasts and JEGIII cells were remarkably similar. In vitro infection resulted in an intracellular viral DNA and hepatitis B surface antigen signal and the secretion of hepatitis B surface antigen into culture medium. The results suggest that both isolated trophoblast cells and trophoblast-derived choriocarcinoma cells are sensitive to infection with HBV in vitro. In addition, we have found that infection of trophoblast cells and JEGIII cells by HBV was enhanced in the presence of tumor necrosis factor alpha. This supports an additional role for tumor necrosis factor alpha in the entry of HBV into trophoblast cells during pregnancy.

Hepatitis C virus (HCV) infection represents a major public health problem throughout the world. The establishment of viral replicons has enhanced our understanding of the mechanism underlying HCV replication. However, the specific virus-host cell interactions involved in HCV RNA replication are not well understood. In the present study, we isolated several human hepatoma Huh-7-derived subclones with a range of HCV RNA replication efficiencies by end-point dilution. Of these, the clones HuhTe4 and HuhTe6 were observed to proliferate at the same rate; however, HuhTe6 supported a significantly greater degree of viral RNA replication. Using cDNA microarray analysis, a total of 36 genes (0.4%) demonstrated variable expression, with a ≧2- fold difference in expression noted between HuhTe4 and HuhTe6. Among genes that are implicated in a variety of functional categories, a subset of these differentially-expressed genes has a role in signal transduction and cell communication, including thioredoxin-interacting protein, Rab6B, sorting nexin 16 and UDPgalactose:ceramide glycosyltransferase. The genes identified in this study should be examined further to determine their roles in HCV RNA replication. The Huh-7 subclones identified in this study provide a tool for identifying novel host factors involved in viral replication.

This study was designed to determine the prevalence of healthy Haemophilus influenzae carriers in a random sample of the preschool population in Kayseri, Turkey. The lack of H. influenzae type b (Hib) disease surveillance and epidemiological data on the throat carriage of Turkish children has caused a delay in the introduction of conjugated Hib vaccination into proposed national vaccination programs. Oropharyngeal cultures were collected and cultured on chocolate agar supplemented with 260 μg/ml bacitracin from 683 children between May and June, 2006. One hundred seven (15.6%) of the 683 children studied were found to be as H. influenzae carriers, and 29 (4.2%) isolates were serotype b. Beta-lactamase production was detected in four isolates (3.7%). According to multivariate analysis, the sex of the child and the number of people sharing the same room with the child significantly influenced the odds of carrying H. influenzae. Age, having older siblings, passive smoking, respiratory infection during the last 30 days, number of people in the household, attending kindergarten or a day-care center, and household income were not significant variables. Our results suggest that there is a strong relationship between exposure to large numbers of children and H. influenzae carriage.

The rate of the onset of methicillin-resistant Staphylococcus epidermidis infections is increasing in Tunisia. We have isolated 32 S. epidermidis strains from dialysis fluid and needle cultures in dialysis service. The strains were identified by classic methods (colonial morphology, Gram staining, catalase test, coagulase test, and DNase test) as well as by API ID32 Staph. Susceptibilities to 18 antibiotics were tested with the ATB Staph kit. Most of the tested strains were resistant to penicillin. In addition, the presence of multidrug resistant strains that showed resistance to different antibiotics was recorded. We have characterized these strains by multiplex PCR assay to identify intercellular adhesion genes icaA/icaD associated with the adhesiveness of staphylococci in biomaterials, and to identify representative resistant genes: oxacillin resistance, mecA; erythromycin methylase (ermA, ermB, and ermC), and macrolide efflux gene (msrA and mef ). The frequency of the carriage of these genes was icaA/icaD (71.9%), mecA (78.1%), ermA (12.5%), ermB (31.3%), ermC (53.1%), msrA (68.8%), and mef (O%). Although the carriage of the genes and the results of susceptibility testing did not match exactly, it could be judged that the PCR identification of antibiotic resistance genes is rapid and supplementary methods for identifying staphylococci or epidemiological study used for the control of nosocomial infection.

Trichophyton tonsurans has been reported to be the causative agent of an epidemic of tinea corporis and tinea capitis among Japanese judoists and wrestlers. A molecular method using restriction enzyme analysis of PCR-amplified fragments targeting the non-transcribed spacer (NTS) region of ribosomal RNA genes in fungal nuclei was applied to a total of 232 strains of T. tonsurans isolated in Japan. Six molecular types, i.e., NTS types I, II, III, IV, V, and VI, were clearly detected in restriction analysis of fragments digested with MvaI and AvaI together. Of the 232 strains, 199 were classified as NTS I, 21 as NTS II, 7 as NTS III, 3 as NTS IV, 1 as type V, and 1 as type VI. Whereas the NTS I strains were found nationwide, most of the NTS II and NTS III strains were limited to central Japan. Of 164 strains isolated from judoists, 160 were classified as NTS I, which suggests that the majority of the cases were caused by a clonal lineage. On the other hand, the 48 strains isolated from wrestlers showed more variety, with 27 strains classified as NTS I, 17 as NTS II, and 4 as NTS III. We concluded that the epidemic was caused by at least three lineages of T. tonsurans. NTS VI strains, the major molecular type among sporadically isolated strains, were not observed among the epidemic strains, and strains of this type did not contribute to the present epidemic.

Ethiopia is one of the countries in which sexually transmitted infections are highly prevalent. However, the data needed to present a realistic picture of the infections are lacking. This study was therefore designed to determine the seroprevalence of syphilis and HIV-1 among pregnant women at the University of Gondar Teaching Hospital. A prospective cross-sectional hospital-based study was conducted between March and June, 2005. Blood samples were collected from 480 pregnant women attending the antenatal clinic of the hospital. Sera were tested for syphilis using the Rapid Plasma Regain (RPR) and Treponema pallidum hemagglutination (TPHA) kits, and serostatus for HIV infection was checked using rapid HIV diagnostic test kits following the manufacturers’ instructions. The mean (±SD) age of the study participants was 26.1 (±7.2) years. The seroprevalence of syphilis was 1%. Antibodies against HIV-1 were detected in 9.6% of the pregnant women. A higher HIV-1 prevalence (13%) was observed in the 25- to 29-year-old age group followed by the 30- to 34-year-old age group (10.2%). Only one subject (2.2%) was found to be positive for both HIV-1 and syphilis. The data indicated a relatively declined prevalence of syphilis and HIV-1 among pregnant women in an urban antenatal clinic. However, incidence and behavioral studies are required to substantiate the findings.

Recent evidence suggests that certain Fc gamma receptor (FcγR) alleles are genetic risk factors for infectious diseases. In this study, we evaluated FcγRIIa polymorphism in patients with brucellosis. In a case-control study, the frequency of two alleles and three genotypes for FcγRIIa were measured by PCR in 150 patients with brucellosis and 125 healthy controls. The H131 and R131 alleles were found in 133 (44.3%) and 167 patients (47.6%), respectively. The frequencies for the three genotypes (a/a, a/r, r/r) were 10 (6.7%), 113 (75.3%) and 27 (18%), respectively. There was no significant difference in the distribution of FcγRIIa genotypes and the two allelic forms between the patients and controls. Our study indicates that FcγRIIa polymorphism is not decisive for the acquisition of brucellosis.

In spite of the routine 2-dose vaccination and three recent supplemental immunizations, Mongolia experienced a measles outbreak in 2001, the largest epidemic in the country since 1984. The majority of cases were reported in the capital city, and the disease incidence was higher in infants and adolescents than in other age groups. Young adults who received the immunization only once may have low immunity, and may be exposed to the virus most frequently. Immunization strategies such as the age range that is targeted for vaccination and the interval between supplemental immunizations should be based on reasonable epidemiological observations.

This report describes a patient presenting mitral native endocarditis due to Campylobacter fetus subsp. fetus, which was revealed by syncope and identified using 16S ribosomal RNA gene sequencing. This gene sequencing method has become the preferred approach to identifying the new emerging pathogens when discrepancies exist between phenotypical tests.

Using the newly designed mismatch amplification mutation assay (MAMA) PCR, we demonstrated the high frequency of amantadine-resistant influenza A (H3N2) viruses isolated during the 2005 - 2006 season by detecting the mutation at amino acid position 31 of the M2 protein (S31N). Further, phylogenetic analyses of the HA1 sequences of the S31N viruses revealed that they comprised a clonal lineage that would result in the common characteristic amino acid changes at positions 193 (Ser to Phe) and 225 (Asp to Asn) of the HA protein. We also demonstrated that the S31N/S193F/D225N viruses had already emerged in Aichi Prefecture by the end of the previous 2004 - 2005 season.

The purpose of this study was to examine the extent of dengue infection in Brunei and to determine the predominant serotype circulating in the country. The study generated useful epidemiological data on dengue infection in Brunei. A total of 271 samples from patients suspected of having dengue infections were selected and analyzed. All patients were seen in clinics and hospitals in Brunei. The samples were collected from April 2005 to April 2006 and transported to the WHO Collaborating Centre for Arbovirus Reference and Research, University of Malaya, Malaysia. The following tests were used to achieve the objectives: in-house IgM-capture enzyme-linked immunosorbent assay, virus isolation in mosquito albopictus cell line (C6/36), and viral RNA detection and serotyping by reverse transcriptase-polymerase chain reaction (RT-PCR). The results show that 45 people were positive for dengue-specific IgM (27 males and 18 females), while RT-PCR detected dengue viral RNA in 12 patients, 3 identified as DEN-1 and 9 as DEN-2. Dengue virus was isolated from 6 patients using the C6/36 cell line; 3 were DEN-2 isolates and 3 were DEN-1 isolates. These data show that dengue virus is circulating in Brunei and the predominant infecting serotype for that period was DEN-2 followed by DEN-1. This study is the first to report the detection and isolation of dengue virus from Brunei using RT-PCR and culture in the C6/36 albopictus mosquito cell line.

The prevalence and pattern of parasitic infestation among 80 HIV/AIDS patients with diarrhea in Madurai, south India, was studied by microscopy. Eighty HIV-negative patients were used as controls. Intestinal parasites were detected in 31 HIV/AIDS patients (38.7%) and in 14 (17.5%) HIV-negative patients, a difference that was statistically significant (P < 0.05). In HIV/AIDS patients with diarrhea, protozoa accounted for the majority of diarrhea cases (Entamoeba spp. 37.5%, Cryptosporidium parvum 28.7%). It is therefore suggested that enteric infections are more common in HIV-infected patients than in HIV-negative persons in south India, and this may be due to differences in immunological profile, susceptibility as well as factors related to sanitation and the environment.

The nature and frequency of mutations in the rpoB gene of rifampicin (RIF)-resistant Mycobacterium tuberculosis clinical isolates vary considerably according to the geographical location, and very little information is available regarding specific mutational patterns in our country. The main objective of this study was to determine the frequency of mutations in the hypervariable region of the rpoB gene in RIF-resistant M. tuberculosis isolates recovered from tuberculosis patients in our region by using the INNO-LiPA Rif. TB kit and to evaluate the performance of the kit for the detection of RIF-resistance. Mutations associated with RIF resistance were studied by line probe assay (LiPA) in 65 RIF-resistant and 56 RIF-susceptible M. tuberculosis strains isolated from different patients in the Aegean region of Turkey. The LiPA identified all susceptible strains (100%) as RIF-sensitive and 63 of 65 (96.9%) phenotypically documented RIF-resistant M. tuberculosis isolates as RIF-resistant, with specific detection of mutation in 44 (67.7%) isolates, whilst 2 strains were identified as RIF-susceptible. The R5-pattern (Ser-531-Leu mutation) was the most frequently observed (35 of 65, 53.8%), followed by the ∆S2-pattern (7.7%) and ∆S4-pattern (7.7%).

Duopath Legionella is a new immunochromatographic assay for the identification of Legionella pneumophila and Legionella spp. As excellent specificity has been previously reported for this kit, we attempted an evaluation of its sensitivity using L. pneumophila serogroup 1 and Legionella anisa strains. Bacterial suspensions of L. pneumophila at concentrations of 1.2×10⁸ cfu/ml and 1.2×10⁷ cfu/ml were detected, but those below 1.2×10⁶ cfu/ml were not recognized by the Duopath kit. After centrifugation and the sediment resuspension, a 2.8×10⁷ cfu/ml concentration of bacterial suspension showed a positive result, but negative results were obtained below 2.8×10⁶ cfu/ml. Also, bacterial suspension with concentrations of 3.2×10⁹ cfu/ml and 1.4×10⁹ cfu/ml after centrifugation of L. anisa were detected, but those below 3.2 × 10⁸ cfu/ml and 1.4 × 10⁸ cfu/ml after centrifugation were not recognized by the Duopath kit. Meanwhile, this kit was less sensitive to the L. pneumophila serogroup 1 suspension, and was more sensitive to the L. pneumophila serogroup 2 and L. anisa than the Binax NOW immunochromatographic kit. It was realized that the sensitivity of this kit is too low for determining the presence of Legionella in water samples. Although this kit may have excellent specificity, it has low sensitivity.

To detect campylobacteriosis and determine the drug susceptibility of causative organisms, we acquired 500 diarrheic samples in Cary-Blair transfer medium from two pediatric hospitals in Tehran between October 2004 and October 2005. The samples were also enriched in Preston broth (with supplements) and defibrinated sheep blood (7%). They were plated from both media on Brucella agar containing antibiotics and blood. Isolates were identified through biochemical tests and by the polymerase chain reaction method. Drug susceptibility testing was performed using the disk diffusion method. In total, 40 Campylobacter strains were isolated (8%). C. jejuni was the dominant species (85.8%) followed by C. coli (14.2%). The rates of resistance to antimicrobial agents were as follows: ciprofloxacin (61.7%), ceftazidime (47%), carbenicillin (35%), tetracycline (20.5%), cefotaxime (14.7%), ampicillin (11.7%), neomycin, erythromycin and chloramphenicol (2.9%), gentamicin, streptomycin, imipenem and colistin (0%). Campylobacter is an important cause of diarrhea among Iranian children. The detection of Campylobacter increases by 25% if samples are treated in enrichment broth prior to plating. The high rate of resistance to ciprofloxacin is alarming, and further investigation into the possible reasons for this is imperative.

Reactive arthritis (ReA) is a sterile arthritis triggered by distal mucosal infection, which suggests a contribution from bacterial products. The pathogenesis of ReA is unclear. There are no international standards for the serological methods used to confirm ReA. In the present work, we analyzed the predominant bacterial component that triggered an immune response in a 24-year-old woman with acute ReA. The candidate bacterial trigger was investigated by measuring the antibacterial antibodies (all immunoglobulin classes and IgA) to Salmonella enteritidis, Shigella flexneri and Yersinia enterocolitica. ELISA for Salmonella gave a positive result. To identify the bacterial component triggering ReA, antibodies to crude lysate, outer membrane proteins (OMP), cytosolic fraction, supernatant proteins and lipopolysaccharide of S. enteritidis were analyzed in sera and synovial fluid (SF) by ELISA, dot blot and Western blot. Among the antigen preparations, the antibody response to OMP was dominant in both serum and SF; a strong reaction to seven OMP bands (50 - 21 kDa) was observed. We concluded that OMP were the main bacterial antigens that trigged ReA in the reported case. Determining the triggering bacterial components in each case can help elucidate the precise causes of ReA and will contribute to the designing of a specific serological diagnostic method for this arthritis.

There is massive enteric bacterial colonization in the periurethral region during infancy. Fecal soiling is considered to be responsible for this colonization. We hypothesized that baby wet wipes containing chemical cleansing compounds, which are used for the cleaning of babies after diaper soiling, could be a contributing factor in this colonization. Thus, the effect on periurethral flora of two different methods of baby cleaning was compared. Periurethral culture samples were obtained from 173 infants who were cleaned with baby wet wipes (Group A, n = 96) or water and napkins (Group B, n = 77) after diaper soiling. The colonization of uropathogens and the presence of flora were analyzed. The results of the periurethral cultures were similar in both groups. The rates of uropathogen colonization only, uropathogen and skin flora colonization, and skin flora only or no growth in Groups A and B were 18.7, 61.5, and 19.8%, and 14.3, 66.2, and 19.5, respectively. The differences between the groups were statistically insignificant (P > 0.05); however, there was no significant difference in the frequency of uropathogen isolation between males and females. We therefore concluded that baby wet wipes are as safe as water for the cleaning of babies after diaper soiling.

Ampicillin-sulbactam (A/S) and amoxicillin-clavulanic acid (AUG) are thought to be equally efficacious clinically against the Enterobacteriaceae family. In this study, the in vitro activities of the A/S and AUG were evaluated and compared against Escherichia coli and Klebsiella spp. Antimicrobial susceptibility tests were performed by standard agar dilution and disc diffusion techniques according to the Clinical and Laboratory Standards Institute (CLSI). During the study period, 973 strains were isolated. Of the 973 bacteria isolated, 823 were E. coli and 150 Klebsiella spp. More organisms were found to be susceptible to AUG than A/S, regardless of the susceptibility testing methodology. The agar dilution results of the isolates that were found to be sensitive or resistant were also compatible with the disc diffusion results. However, some differences were seen in the agar dilution results of some isolates that were found to be intermediately resistant with disc diffusion. In E. coli isolates, 17 of the 76 AUG intermediately resistant isolates (by disc diffusion), and 17 of the 63 A/S intermediately resistant isolates (by disc diffusion) showed different resistant patterns by agar dilution. When the CLSI breakpoint criteria are applied it should be considered that AUG and A/S sensitivity in E. coli and Klebsiella spp. Strains may show differences.

An indirect immunofluorescent assay to detect antibodies against the lipopolysaccharide (LPS) of Burkholderia pseudomallei and taxonomically closely related species was developed with the Luminex system. LPSs of Pseudomonas aeruginosa, Burkholderia cepacia, Burkholderia thailandensis, Burkholderia vietnamiensis, B. pseudomallei, and Burkholderia mallei were successfully conjugated to Luminex microspheres. Antibodies measured against the LPS of B. pseudomallei-conjugated Luminex beads only cross-reacted with those of two genetically closely related species, B. mallei and B. thailandensis (previously classified as non-pathogenic arabinose-negative B. pseudomallei). However, this system could distinguish other closely related species from B. pseudomallei. This assay is able to detect significantly high levels of anti-LPS antibodies of B. pseudomallei in serum from patients with culture-proven melioidosis.