Acute tick-borne encephalitis during pregnancy - An alarming combination.

We report on a pregnant patient who contracted tick-borne encephalitis (TBE) during her second trimester in an endemic region in Southern Germany. The patient presented with typical symptoms including fever and headache, and TBE infection was confirmed by positive blood and cerebrospinal fluid (CSF) testing. During acute infection there was no evidence of pregnancy complications, and the mother recovered well. We performed a clinical follow-up examination of both mother and child eight months after the diagnosis of TBE, which revealed no signs of sequelae. This case study presents rare evidence of TBE infection during pregnancy and may provide guidance for both physicians as well as mothers-to-be dealing with TBE.

Transfusion-transmitted hepatitis E in Germany, 2013.

The reported IgG seroprevalence against hepatitis E virus (HEV) in German blood donations is 6.8%, and HEV RNA detected in 0.08%, but documented evidence for HEV transmission is lacking. We identified two donations from a single donor containing 120 IU HEV RNA/mL plasma and 490 IU/mL. An infectious dose of 7,056 IU HEV RNA was transmitted via apheresis platelets to an immunosuppressed patient who developed chronic HEV. Further, transmission was probable in an immunocompetent child.

Performance of a novel microarray multiplex PCR for the detection of 23 respiratory pathogens (SYMP-ARI study).

Symptoms of acute febrile respiratory tract infection are often unspecific, but the rapid identification of pathogens allows optimised patient management. The objective of this study was to evaluate a novel multiplex polymerase chain reaction (PCR) suspension microarray which detects 19 viral and four atypical bacterial targets. A comprehensive set of sensitive monoplex real-time PCR assays was used for each pathogen as the gold standard. A panel of archived as well as 300 prospectively collected clinical samples was analysed by both methods. At least one target was detected in 165/300 (55 %) samples by monoplex PCR and in 140/300 (46 %) samples by multiplex PCR, respectively. The positivity rate was significantly higher in paediatric patients compared to adults [126/154 (82 %) vs. 39/146 (27 %) by monoplex and 114/154 (74 %) vs. 26/146 (18 %) by multiplex PCR, respectively]. Among all samples, 17/300 (5.6 %) were positive for atypical bacteria by monoplex and 8/300 (2.6 %) by multiplex PCR, respectively. Multiple detections were recorded in 35/300 (11.6 %) samples by monoplex and 26/300 (8.7 %) by multiplex PCR. For the most common pathogens, the sensitivity ranged from 57 to 93 % and the specificity ranged from 95 to 100 %. The overall concordance between both methods was 77 % [95 % confidence interval (CI) 72-81 %]. False-negative results by multiplex PCR were mainly due to the low target concentration. Compared to monoplex PCR, the novel microarray assay proved its principle but displayed overall lower sensitivities, potentially restricting its use to paediatric patients. For some targets, only small numbers of positive samples were available, requiring larger studies to firmly assess the sensitivity and specificity.

Detection of influenza A(H1N1)v virus by real-time RT-PCR.

Influenza A(H1N1)v virus was first identified in April 2009. A novel real-time RT-PCR for influenza A(H1N1)v virus was set up ad hoc and validated following industry-standard criteria. The lower limit of detection of the assay was 384 copies of viral RNA per ml of viral transport medium (95% confidence interval: 273-876 RNA copies/ml). Specificity was 100% as assessed on a panel of reference samples including seasonal human influenza A virus H1N1 and H3N2, highly pathogenic avian influenza A virus H5N1 and porcine influenza A virus H1N1, H1N2 and H3N2 samples. The real-time RT-PCR assay for the influenza A matrix gene recommended in 2007 by the World Health Organization was modified to work under the same reaction conditions as the influenza A(H1N1)v virus-specific test. Both assays were equally sensitive. Clinical applicability of both assays was demonstrated by screening of almost 2,000 suspected influenza (H1N1)v specimens, which included samples from the first cases of pandemic H1N1 influenza imported to Germany. Measuring influenza A(H1N1)v virus concentrations in 144 laboratory-confirmed samples yielded a median of 4.6 log RNA copies/ml. The new methodology proved its principle and might assist public health laboratories in the upcoming influenza pandemic.

Immunization of liver and renal transplant recipients: a seroepidemiological and sociodemographic survey.

Several life-threatening infections, a major risk to adult solid organ transplant (SOT) recipients on immunosuppressive therapy, can be prevented by immunization. We analyzed sociodemographic parameters and the immunization status of adult liver transplant recipients (LTX-R, n=267) and renal transplant recipients (RTX-R, n=197) SOT recipients at the Transplantation Center, Berlin, Germany. Date, number, and provider of recommended vaccines were recorded and seroprotection rates determined. The social status in both groups was similar. Most patients (89%) were not adequately informed about immunizations; and if informed, main sources were physicians (47%) and the media (40%). Vaccinations were predominantly provided by family doctors (LTX-R, 66%; RTX-R, 31%) or hemodialysis centers (RTX-R, 37%). Before transplantation, RTX-R had significantly more often received booster vaccinations against tetanus and diphtheria (P<0.005), and a primary hepatitis B immunization (55%); whereas in LTX-R, post-transplant vaccinations against hepatitis A (16%) and pneumococcal disease (13%) were more frequent. Seroprotection rates against tetanus were fairly high in LTX-R (85.3%) and RTX-R (86.8%), and considerably lower for diphtheria, hepatitis A, and influenza. Immunization rates are too low in SOT recipients. Improvement will depend on a more active role of health care providers.

Performance of a polymer-based DNA chip platform in detection and genotyping of human papillomavirus in clinical samples.

Human papillomavirus (HPV) plays a key role in the development of cervical and laryngeal cancers. The aim of our study was to compare the performance of a new hydrogel-based HPV genotyping biochip assay (Biochip) to a commercially available and CE-marked conventional PCR followed by reverse hybridization (GenID-PCR). One hundred twenty-three samples were available for the study. Of these samples, 101/123 were gynecological swabs, 8/123 were swabs or biopsy samples of genital warts, 7/123 were biopsy samples of otorhinolaryngeal lesions, 5/123 were samples of skin warts, and 2/123 were samples of orolabial abnormalities. These molecular methods for HPV genotyping showed comparable sensitivity and specificity. However, 19/123 of the results were discrepant. Specifically, Biochip showed better performance in the detection of multiple infections, especially when more than one high-risk genotype was present. Due to the different probe configurations used in the two assays, GenID-PCR achieves only group-specific detection of many HPV genotypes, whereas Biochip allows for specific identification. Overall, the newly developed HPV chip system (Biochip) proved to be a suitable tool for HPV detection and genotyping; it also proved to be superior for establishing HPV genotyping methods.

Comparison of nine commercially available assays for quantification of antibody response to hepatitis B virus surface antigen.

Quantitative measurement of anti-HBs is used to evaluate the response to hepatitis B vaccination in health care workers and to optimize postexposure management. The different guidelines for hepatitis B vaccination and booster policy imply that the measurement of anti-HBs levels by different assays is accurate and consistent, yielding comparable quantitative results. We measured anti-HBs levels in 200 serum samples from patients and health care professionals by nine different anti-HBs assays and compared the quantitative results and the performance characteristics of the different test systems. The assay specificity ranged between 96.8 and 100% when sera from individuals without a vaccination history and with negative anti-HBc status were defined as true negatives. Sensitivity ranged between 93.5 and 100%. A high number of sera showed discrepancies between measurements by the different systems. The mean coefficient of variation between the different measurements was 47.1% (range, 15.0 to 201.0%), and the factors of multiplication ranged from 2.8 to 105. Hemolysis or lipemia did not seem to influence the measurement, and there was no difference between anti-HBc-positive and -negative individuals. The classical enzyme immunoassays tend to find lower anti-HBs levels than the automated systems, with higher values by the Abbott AXSYM assay. The serial dilution of the international standard preparation was measured accurately by most of the assays. In conclusion, the quantitative measurement of anti-HBs levels is not reliable, even though an international standard is used for the calibration of the systems. Some systems showed specific problems that should be addressed by the manufacturers.

Active vaccination in patients with common variable immunodeficiency (CVID).

Active vaccination of CVID patients with standard vaccines has rarely been studied in depth although some patients have been shown to develop transient vaccine-specific immunity. We addressed the question whether these patients can be identified by functional classification of their B cell subsets in vitro. Twenty-one CVID patients receiving regular IgG substitution were immunized with anti-peptide and anti-polysaccharide vaccines. Humoral vaccination responses were compared to the numbers of circulating memory B cells, CD21(low) B cells and the capacity to produce antibodies in vitro. Our findings allow four conclusions: (1) positive vaccination responses are not contradictory to the diagnosis of CVID; they occurred against polypeptide vaccines in 23% and against polysaccharide antigens in 18% of all vaccinations. (2) Class-switched antibody responses occur preferentially in patients of CVID group II. (3) A normal percentage of IgM memory B cells is necessary but not sufficient for a vaccination response to polysaccharide antigens. (4) Active vaccination in addition to IgG replacement therapy should be performed in patients of CVID type II - especially in case of vaccines for which passive protection cannot be guaranteed.

Demand for evaluation of vaccination antibody titers in children considered for renal transplantation.

Vaccinations are recommended for achieving protection against vaccine-preventable infections in solid-organ transplant recipients. In order to evaluate the protection at the time of renal transplantation, the antibody titers against measles, mumps, rubella, varicella, hepatitis B, diphtheria, and tetanus were determined in 35 children one month prior to transplantation. Only 26% of patients on dialysis listed for transplantation showed protective antibodies against all tested pathogens. Particularly, low protection was found for hepatitis B. Children younger than four yr showed significantly lower protective antibody titers compared with older children for almost all vaccines. Children who completed vaccination in the last six months to six yr prior to renal transplantation showed higher rates of protective antibody titers against all pathogens compared with children who had vaccination more than six yr before transplantation. Preventive strategies in children with chronic renal failure include repeated measurements of serum antibodies and appropriate revaccination if titers decline. Our results underline the demand for continuous surveillance of specific antibody titers against vaccine-preventable diseases in the risk group of renal transplant recipients.

[Evaluation of an enzyme immunoassay for the determination of the status of immunity against diphtheria before and after vaccination]. / Validierung eines Enzymimmunoassays zur Bestimmung des Diphtherieimmunstatus vor und nach Impfung.

PURPOSE: Assessment of an Easy-to-Perform Immunoenzymatic Assay for Detecting Diphtheria Immunity in the Population. Can an immunoassay replace or support the toxin neutralisation test? SERA AND METHODS: Sera of 75 adults were collected before and after vaccination and tested in the vero cell neutralisation test and the immunoassay. RESULTS: The sensitivity of the EIA was 73 and 94%, specificity was 74 and 67% before and after vaccination, respectively. Positive or negative predictive value was 70 and 96% or 78 and 49% before and after vaccination, respectively. The correlation coefficient changed from 0.43 to 0.77 after vaccination. A comparison of the quantitative results of the test showed differences up to 19 times in individual cases. CONCLUSIONS: Only cases with relatively high antibody titres (e.g. after booster vaccination) show EIA high sensitivity. Linear correlation is also high in such cases. If you expect low antitoxin levels as is the case before booster vaccination, EIA will then often be false positive or negative. Such studies should be limited to the neutralisation method.

Hepatitis G virus RNA and hepatitis G virus antibodies in renal transplant recipients: prevalence and risk factors.

BACKGROUND: Hepatitis G virus (HGV/GBV-C) RNA indicating current infection has been frequently isolated from the sera of transplant recipients and other multitransfused individuals. Lifetime exposure to the virus, however, is unknown. We carried out a study to determine the prevalence and risk factors of HGV antibodies and of HGV RNA among renal transplant recipients, and to investigate possible associations between HGV RNA and immunosuppressive treatment. METHODS: HGV RNA was detected by reverse transcriptase-polymerase chain reaction, and HGV antibodies (anti-E2) by a newly developed immunoassay. To assess risk factors for HGV exposure, univariate and multivariate analysis was performed. RESULTS: Of the 221 patients, 14% were HGV RNA positive and 40% had HGV antibodies. Both HGV RNA and anti-HGV were present in only two individuals. Thus, the overall HGV exposure prevalence was 53%. It increased significantly with the number of blood transfusions. In logistic regression, the adjusted HGV exposure prevalence odds ratio was 5.7 (95% confidence interval [CI]: 2.2-15) among patients with > or =10 transfusions (baseline: no transfusions). Other independent risk factors were a longer duration of hemodialysis and a longer time interval since transplantation. HGV viremia was not associated with the type of immunosuppressive treatment. Alanine aminotransferase levels were not significantly increased among HGV RNA-positive patients. CONCLUSIONS: Much higher proportions of renal transplant recipients were exposed to HGV than is suggested by HGV RNA detection alone. The majority of infected individuals apparently eliminate the virus over time. Contaminated blood transfusions have to be regarded as a main risk factor for HGV infection.

Tumor necrosis factor alpha and interleukin 6 release induced by antibiotic killing of Pseudomonas aeruginosa and Staphylococcus aureus.

Using a recently described ex vivo model, tumor necrosis factor-alpha and interleukin-6 released by peripheral blood monocytes after killing of Pseudomonas aeruginosa and Staphylococcus aureus by ceftazidime, imipenem, and meropenem was measured. Cytokine release was highest with ceftazidime and lowest with imipenem for both bacteria (p < 0.05), although cytokine concentrations were much lower after killing of Staphylococcus aureus. Differences in cytokine release rates induced by various cell-wall active antibiotics have not yet been described for gram-positive organisms and should be studied further.

Routine immunizations in adult renal transplant recipients.

BACKGROUND: Vaccination guidelines for transplant recipients include regular boosters of tetanus, diphtheria, and inactivated polio vaccine, but there are few published data on the efficacy of these vaccines in patients receiving immunosuppressive therapy. METHODS: Serum antibody values were evaluated before and 4 weeks after tetanus, diphtheria, and inactivated polio vaccination in 164 renal transplant recipients compared with healthy controls. Twelve months later, antibody levels were evaluated in 55 patients. RESULTS: Prebooster tetanus antitoxin values were lower in transplant recipients than in controls. All patients developed protective tetanus antibody levels (> or = 0.01 IU/ml) after vaccination. After 12 months, serum antibodies had decreased, but all patients maintained protective values. Diphtheria antitoxin titers before and after booster vaccination were lower in patients than in controls: 88.5% of patients and 96.2% of controls developed protective diphtheria antibody values. Twelve months after vaccination, diphtheria antitoxin values were below the protective level (0.1 IU/ml) in 38% of patients. Prebooster antibody values to poliovirus types 1 and 3 were comparable in patients and controls, whereas antibodies to poliovirus type 2 were lower in transplant recipients. Seroprotection rates and geometric mean antibody titers after vaccination were equivalent between the two groups for all three poliovirus types. No difference was observed in antibody levels between patients on different immunosuppressive drug regimens. Adverse reactions were significantly less often reported by transplant recipients. CONCLUSIONS: In transplant recipients, tetanus and inactivated polio vaccinations are well tolerated and induce protective antibody levels; diphtheria vaccination as currently recommended is less effective and protective antitoxin values decrease rapidly in these patients within 1 year after vaccination.

[Hospital infections in gynecology and obstetrics. An inclusive prevalence study in Germany]. / Krankenhausinfektionen in Gynäkologie und Geburtshilfe. Eine bundesweite Prävalenzstudie (NIDEP).

In a German multicenter survey, 2206 gynaecological patients in 72 randomly selected hospitals were examined for the prevalence of nosocomial infections and possible risk factors. Hospital-acquired infections were diagnosed in 1.45% of the patients. The most common localisation was the urinary tract (0.91%). Septicaemia, vaginitis and infections of the upper and lower airways were only rarely seen. The following endogenous risk factors were identified: diseases of the cardiovascular system (16.1%), malignancies (12.2%) preexisting infections (6.1%), obesity (5.9%), and diabetes (5.0%). The most common exogenous risk factors were peripheral venous catheters (19.9%), catheterisation of the urinary tract (7.2%) and wound drainage (28.6%). 49% of the patients who underwent caesarean section and 50% of the hysterectomy patients received antimicrobial chemoprophylaxis.

Malaria Chemoprophylaxis in German Tourists: A Prospective Study on Compliance and Adverse Reactions.

Background: The number of tourists visiting malaria endemic regions is continuously increasing. The risk of aquiring malaria infection can largely be prevented by the regular use of chemoprophylactic drugs combined with using protective measures against mosquito bites. In a prospective study we wished to determine the tolerability of chemoprophylactic drugs and the factors that influence compliance with malaria chemoprophylaxis and antimosquito measures. Method: German travelers (n = 6504) who attended the Berlin Institute of Tropical Medicine in Berlin for pretravel medical advice were interviewed by phone 4 weeks after their journeys about compliance with the recommended malaria chemoprophylaxis and the incidence of side effects. Results: Compliance was better with mefloquine (94.5%) than with chloroquine (85.9%) (p<.001) or chloroquine plus proguanil (79.8%) (p<.001). Compliance was influenced by the purpose of travel, duration of stay, places of stay, and adverse reactions. Side effects occurred in 20.6% of the travelers who took chemoprophylactic drugs. There was no significant difference in the incidence of side effects between the three drug regimens, but people who took mefloquine more often reported neuropsychiatric reactions (6.5% versus 3.9% with chloroquine and 3.6% with chloroquine and proguanil; p<.001)). Side effects were usually mild to moderate and in no instance required hospitalization. People who took their drugs with meals less often reported side effects (15.2%) (p<.01). Conclusion: The knowledge of user profiles (and particular factors that presage side effects and noncompliance) may help us to improve pretravel counseling, thereby reducing the risk that travelers may acquire malaria infection.