It has been proposed that patients who develop Clostridium difficile-associated disease (CDAD) do so because they are unable to mount an adequate immune response. Serum was collected from three groups of elderly in-patients: (i) cases (n521) of CDAD, being toxin A/B-positive; (ii) carriers (n521) asymptomatic for CDAD (no diarrhoea) but at least toxin or culture positive; and (iii) controls (n526) asymptomatic for CDAD and negative for both C. difficile toxin and culture. The age and gender of each group were compared, and the colonizing strains were ribotyped and toxinotyped. Serum antibodies (IgG and IgM) were measured by ELISA using different antigen preparations: EDTA extract (containing cell-surface proteins and carbohydrates), guanidine hydrochloride extract (surface-layer proteins), aqueous phenol-extracted lipocarbohydrate (LC); crude toxin (dialysis culture supernatant) and purified toxin A. LPS from Escherichia coli was used as a control antigen. Antibodies were also tested for toxin neutralization on tissue monolayers and for binding to EDTA-extracted antigens by Western blotting. IgG antibody measurements to cytomegalovirus (CMV) were included as an indicator of potential immunosenescence. Results showed that the patient groups were well matched by age and gender, and the colonizing strains were similar in cases and carriers, being predominantly ribotype 001 and toxinotype 0. By ELISA, IgG levels to most of the antigens were highest in the cases and lowest in the controls, with the exception of antibodies to the LC, which were higher in the controls than the cases. Levels in the carriers tended to be of intermediate level or similar to the controls. For all antigens, the levels of IgM were not significantly different among cases, carriers and controls. Serum from all groups was able to neutralize the cytotoxic action of toxin on both Vero and Caco2 cells, and all to a similar extent. Western blots showed an overall higher level of IgG antibodies to the EDTA-extracted antigens in the cases. The results of the CMV ELISA showed that specific IgG was detected in more cases (78 %) than carriers and controls (both 65 %), but this difference in seropositivity was not significant. The conclusion is that, during symptomatic infection, patients respond to protein antigens of C. difficile in a manner typical of a secondary antibody response, with no evidence that an inability to respond predisposes to the appearance of symptoms.

Caspase-associated recruitment domain-9 (CARD9) deficiency is an autosomal-recessive primary immunodeficiency with genetic defects in Th17 immunity marked by susceptibility to recurrent and invasive Candida infections. We present a case of relapsing Candida albicans meningoencephalitis over 1-year period despite appropriate antifungal therapy. We detected a homozygous p.Q295X mutation in CARD9 as well as a defective interleukin-17 and interferon gamma synthesis in Enzyme-Linked ImmunoSpot tests. We achieved complete clinical remission, and improvement of interleukin-17 secretion with subcutaneous granulocyte colony-stimulating factor) treatment.

Clostridium difficile isolates (n5149) collected in south-east Scotland between August and October 2005 were typed by four different methods and their susceptibility to seven different antibiotics was determined. The aims were to define the types of strain occurring in this region and to determine whether there were any clonal relationships among them with respect to genotype and antibiotic resistance pattern. Ribotyping revealed that 001 was the most common type (n5113, 75.8 %), followed by ribotype 106 (12 isolates, 8.1 %). The majority of the isolates (96.6 %, n5144) were of toxinotype 0, with two toxinotype V isolates and single isolates of toxinotypes I, IV and XIII. PCR and restriction analysis of the fliC gene from 147 isolates gave two restriction patterns: 145 of pattern VII and two of pattern I. Binary toxin genes were detected in only three isolates: two isolates of ribotype 126, toxinotype V, and one isolate of ribotype 023, toxinotype IV. S-types showed more variation, with 64.5 % (n540) of the common S-type (4939) and 21 % (n513) of S-type 4741, with six other S-types (one to three isolates each). All ribotype 001 isolates were of the same S-type (4939), with three isolates of other ribotypes being this S-type. No resistance was found to metronidazole or vancomycin, with resistance to tetracycline only found in 4.3 % of the isolates. A high proportion of isolates were resistant to clindamycin (62.9 %), moxifloxacin, ceftriaxone (both 87.1 %) and erythromycin (94.8 %). Resistance to three antibiotics (erythromycin, clindamycin and ceftriaxone) was seen in 66 isolates, with erythromycin, ceftriaxone and moxifloxacin resistance seen in 96 isolates. Resistance to all four of these antibiotics was found in 62 isolates and resistance to five (the above plus tetracycline) in one isolate: a ribotype 001, toxinotype 0 strain. Whilst ribotype 001 was the most commonly encountered type, there was no evidence of clonal relationships when all other typing and antibiotic resistance patterns were taken into account. INTRODUCTIONClostridium difficile is an anaerobic, Gram-positive, sporeforming bacillus. It is commonly associated with a spectrum of disease referred to as C. difficile-associated disease (CDAD), which can range from uncomplicated mild diarrhoea to lethal toxic megacolon and possible colon perforation (Johnson & Gerding, 1998). It is considered to be the leading cause of nosocomially acquired diarrhoea in adults and can be responsible for large outbreaks (Kelly & LaMont, 1998). There is a view that the severity of the disease is increasing. The new hypervirulent type (ribotype 027, toxinotype III, pulse-field NAP1) in North America and several European countries has been associated with more severe and fatal cases (McDonald et al., 2005;Kuijper et al., 2006a;Hubert et al., 2007).As elsewhere, C. difficile is rarely cultured in Scotland and laboratory diagnosis depends on the detection of toxins A and/or B in faeces. Several phenotypic and molecular methods have been applied to determine the relatedne...

BackgroundRecognition of nuclear dense fine speckled (DFS) pattern by indirect immunofluorescence (IIF) is not easy. Thus, confirming the presence of these antibodies might be needed. In this study, we aimed to determine the frequency of DFS pattern in our diagnostic laboratory and to investigate the presence of anti-DFS70 antibodies in samples showing DFS pattern by two commercially available research kits retrospectively.Material and methodsSeventy-four sequential serum samples with DFS pattern on HEp2010 cell substrates by IIF were included in this study. The semiquantitative DFS70 ELISA Kit (MBL International Corporation, Woburn, UK) was used for detection of anti-DFS70 antibodies in these samples. Twenty selected samples were tested for the presence of anti-DFS70 antibodies using ANA Line Immunoassay (LIA) (Immco Diagnostics, New York, USA).ResultsSixty-two (83.8%) of 74 serum samples were found positive with ELISA, when 15 U/ml was taken as a reference value. Among 18 samples that were found positive by ELISA, five were negative for anti-DFS70 antibodies by LIA, while 13 were found positive. The lowest ELISA result of the sample that was positive by LIA was found to be 45.3 U/ml. When 45.3 U/ml was considered as a reference value, 45 (60.8%) of 74 serum samples were positive by ELISA. Nineteen of 20 patients had no SARD, while one had systemic lupus erythematosus (SLE).ConclusionsDFS pattern should be confirmed with an objective method such as ELISA, LIA, or IB. We think that confirmation tests for detection of anti-DFS70 antibodies should be included in diagnostic algorithms.

* Bu çalışmada kullanılan hasta verilerinin bir kısmı, 4. Ulusal Viroloji Kongresi (23-26 Haziran 2011, İstanbul)'nde sunulmuştur. ÖZETPrimer BK virus (BKV) enfeksiyonları genellikle erken çocukluk döneminde kazanılmakta ve asemptomatik olarak geçirilmektedir. Dünyadaki erişkin popülasyonlarda BKV seroprevalansı %90'a kadar ulaşa-bilir. Primer enfeksiyondan sonra virus ürogenital sistemde latent olarak kalmaktadır. Renal transplant alı-cılarında, BKV'nin neden olduğu primer enfeksiyonlar ve reaktivasyonlar, hastaların %10'unu etkileyebilmekte ve önlem alınmadığı taktirde bu hastaların yarısından fazlası BKV nefropatisi (BKVN) nedeniyle böbreklerini kaybetmektedir. BKVN'ye bağlı greft kaybını engellemenin tek yolu, transplantasyon sonrasında BK virus enfeksiyonlarının takibi ve BKVN geliştiren hastaların erken dönemde tanınıp etkin bir biçimde tedavi edilmeleridir. Bu çalışmada, pediatrik renal transplant alıcılarında, transplantasyon sonrası dönemde, idrar ve plazma örneklerinden, gerçek zamanlı polimeraz zincir reaksiyonu (rtPCR) ile BKV enGeliş Tarihi

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DFS pattern detected by IIF and isolated anti DFS70 antibody positivity detected by IB show high concordance. However IIF results should be confirmed because of the patterns that can be misidentified as DFS pattern. The presence of anti-DFS70 antibodies, which help to exclude SARD, prevent further unnecessary referral demands.

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