Performance of T-Track® TB, a Novel Dual Marker RT-qPCR-Based Whole-Blood Test for Improved Detection of Active Tuberculosis.

Tuberculosis (TB) is one of the leading causes of death by an infectious disease. It remains a major health burden worldwide, in part due to misdiagnosis. Therefore, improved diagnostic tests allowing the faster and more reliable diagnosis of patients with active TB are urgently needed. This prospective study examined the performance of the new molecular whole-blood test T-Track® TB, which relies on the combined evaluation of IFNG and CXCL10 mRNA levels, and compared it to that of the QuantiFERON®-TB Gold Plus (QFT-Plus) enzyme-linked immunosorbent assay (ELISA). Diagnostic accuracy and agreement analyses were conducted on the whole blood of 181 active TB patients and 163 non-TB controls. T-Track® TB presented sensitivity of 94.9% and specificity of 93.8% for the detection of active TB vs. non-TB controls. In comparison, the QFT-Plus ELISA showed sensitivity of 84.3%. The sensitivity of T-Track® TB was significantly higher (p < 0.001) than that of QFT-Plus. The overall agreement of T-Track® TB with QFT-Plus to diagnose active TB was 87.9%. Out of 21 samples with discordant results, 19 were correctly classified by T-Track® TB while misclassified by QFT-Plus (T-Track® TB-positive/QFT-Plus-negative), and two samples were misclassified by T-Track® TB while correctly classified by QFT-Plus (T-Track® TB-negative/QFT-Plus-positive). Our results demonstrate the excellent performance of the T-Track® TB molecular assay and its suitability to accurately detect TB infection and discriminate active TB patients from non-infected controls.

Omicron's binding to sotrovimab, casirivimab, imdevimab, CR3022, and sera from previously infected or vaccinated individuals.

SARS-CoV-2 Omicron is the first pandemic variant of concern exhibiting an abrupt accumulation of mutations particularly in the receptor-binding domain that is a critical target of vaccination induced and therapeutic antibodies. Omicron's mutations did only marginally affect the binding of ACE2, and the two antibodies Sotrovimab and CR3022 but strongly impaired the binding of Casirivimab and Imdevimab. Moreover, as compared with Wuhan, there is reduced serum reactivity and a pronounced loss of competitive surrogate virus neutralization (sVN) against Omicron in naïve vaccinees and in COVID-19 convalescents after infection and subsequent vaccination. Finally, although the booster vaccination response conferred higher titers and better sVN, the effect was nonetheless significantly lower compared with responses against Wuhan. Overall, our data suggest that the antigenicity of Omicrons receptor binding motive has largely changed but antibodies such as Sotrovimab targeting other conserved sites maintain binding and therefore hold potential in prophylaxis and treatment of Omicron-induced COVID-19.

Low Adherence to Pneumococcal Vaccination in Lung Cancer Patients in a Tertiary Care University Hospital in Southern Germany.

INTRODUCTION: The aim of this study was to investigate the adherence to vaccinations, especially pneumococcal vaccinations, in lung cancer patients. METHODS: the study was performed at the University Hospital Regensburg, Germany. All patients with a regular appointment scheduled between 1 December 2020 and 29 April 2021 and who provided informed consent were included. Available medical records, vaccination certificates, and a questionnaire were analyzed. RESULTS: we included 136 lung cancer patients (NSCLC n = 113, 83.1%, SCLC n = 23, 16.9%). A correct pneumococcal vaccination according to national recommendations was performed in 9.4% (12/127) of the patients. A correct vaccination was performed for tetanus in 50.4% (66/131), diphtheria in 34.4% (44/128), poliomyelitis in 25.8% (33/128), tick-borne encephalitis in 40.7% (24/59), hepatitis A in 45.5% (7/11), hepatitis B in 38.5% (5/13), shingles in 3.0% (3/101), measles in 50.0% (3/6), pertussis in 47.7% (62/130), influenza in 54.4% (74/136), and meningococcal meningitis in 0% (0/2) of the patients. CONCLUSION: adherence to pneumococcal vaccinations, as well as to other vaccinations, is low in lung cancer patients.

Increased neutralization of SARS-CoV-2 Delta variant after heterologous ChAdOx1 nCoV-19/BNT162b2 versus homologous BNT162b2 vaccination.

Heterologous SARS-CoV-2 vaccine approaches with a second mRNA-based vaccine have been favored in the recommendations of many countries over homologous vector-based ChAdOx1 nCoV-19 vaccination after reports of thromboembolic events and lower efficacy of this regimen. In the middle of 2021, the SARS-CoV-2 Delta variant of concern (VoC) has become predominant in many countries worldwide. Data addressing the neutralization capacity of a heterologous ChAdOx1 nCoV-19/mRNA-based vaccination approach against the Delta VoC in comparison to the widely used homologous mRNA-based vaccine regimen are limited. Here, we compare serological immune responses of a cohort of ChAdOx1 nCoV-19/BNT162b2-vaccinated participants with those of BNT162b2/BNT162b2 vaccinated ones and show that neutralization capacity against the Delta VoC is significantly increased in sera of ChAdOx1 nCoV-19/BNT162b2-vaccinated participants. This overall effect can be attributed to ChAdOx1 nCoV-19/BNT162b2-vaccinated women, especially those with more severe adverse effects leading to sick leave following second immunization.

Human bocavirus 1 respiratory tract reactivations or reinfections in two adults, contributing to neurological deficits and death.

Human bocavirus 1 (HBoV1) of the family Parvoviridae causes mild to life-threatening respiratory tract infections in young children, but, due to widespread immunity, it is uncommon in adults. HBoV1 reinfections or reactivations leading to casualties are rare, but might be underdiagnosed. We report two young adults, one previously healthy and one immunosuppressed, with rare diagnostic patterns of HBoV1 respiratory tract infection. Both patients exhibited very high loads of HBoV1 DNA in respiratory samples. The immunosuppressed patient was also HBoV1 DNA-positive in blood, stool and a colon biopsy, but exhibited prior HBoV1-specific high-avidity IgG and weak IgM positivity 9 months before the respiratory symptoms. Likewise, the previously healthy patient exhibited HBoV1 IgG of high avidity and very weak IgM in serum, pointing to prior immunity, but with a seroconversion in cerebrospinal fluid. This patient also showed strong HBoV2 cross-reactivity. The molecular and serological results, together with their ages, suggest that both patients exhibited unusual reinfection or reactivation of HBoV1, contributing to neurological deficits and death.

Adherence to STIKO recommendations in patients with pulmonary disease in southeast Germany.

BACKGROUND: Vaccinations are among the most effective preventative healthcare measures. The aim of this cross-sectional study was to evaluate the adherence of adults with pre-existing pulmonary conditions to the national vaccination schedule and to identify reasons for poor adherence. METHODS: All patients with an appointment at Donaustauf hospital between October 2019 and April 2020 were asked to bring their vaccination certificates for evaluation and to compete a questionnaire. To determine the adherence vaccination certificates and patients' comorbidities were correlated with the national recommendations of the German Standing Committee on Vaccination (STIKO). RESULTS: 571 (65.6%) of all patients believed that their vaccination status was up-to-date. An appropriate vaccination status according to national recommendations (STIKO) was documented as follows: tetanus 56.4% (375/665), diphtheria 43.2% (292/676), poliomyelitis 28.5% (189/662), tick-borne encephalitis 45.4% (300/659), hepatitis A 31.0% (18/58), hepatitis B 34.6% (27/78), shingles 1.2% (6/489), influenza 21.0% (125/596, season 2019/2020), measles 38.3% (31/81), rubella 33.3% (7/21), pneumococcal disease 29.5% (175/593), pertussis 54.2% (365/674) and haemophilus influenza type b 100% (1/1). Adherence to rabies (0/2), varicella (0/28), meningococcal type ACWY (0/36) and type b (0/36) was 0%. 72% of patients would follow a physician's recommendation to get vaccinated. CONCLUSION: Adherence to STIKO recommendations was poor. However, patients are willing to follow a physician's recommendation for vaccination.

Primary vaccination in adult patients after allogeneic hematopoietic stem cell transplantation - A single center retrospective efficacy analysis.

Allogeneic hematopoietic stem cell transplantation (alloHSCT) results in a loss of humoral immunity and subsequent risk for severe infections. Thus, re-vaccination is required but may fail due to incomplete immune reconstitution. We retrospectively analyzed predictors of immune response to primary vaccination applied according to the EBMT (European Blood and Marrow Transplantation Group) recommendations. Serologic response to vaccination against diphtheria (D), tetanus (T), Bordetella pertussis (aP) and Haemophilus influenzae (Hib) (administrated as combined DTaP-Hib-IPV vaccination) was studied in 84 alloHSCT patients transplanted between 2008 and 2015 (age at alloHSCT: 18.6-70.6 years). All patients with a relapse-free survival of ≥9 months, at least 3 consecutive vaccinations and absence of intravenous immunoglobulin administration within 3 months before and after vaccination met the primary inclusion criteria. Additionally, immunological response to a pneumococcal conjugate vaccine was analyzed in a subgroup of 67 patients. Patients' characteristics at the time of first vaccination were recorded. Responses were measured as vaccine-specific antibody titers. Regarding DTaP-Hib-IPV vaccination, 89.3% (n = 75) of all patients achieved protective titers to at least 3 of the 4 vaccine components and were thus considered responders. 10.7% (n = 9) of the patients were classified as non-responders with positive immune response to less than 3 components. Highest response was observed for Hib (97.4%), tetanus (95.2%) and pneumococcal vaccination (83.6%) while only 68.3% responded to vaccination against Bordetella pertussis. Significant risk factors for failure of vaccination response included low B cell counts (p < 0.001; cut-off: 0.05 B cells/nl) and low IgG levels (p = 0.026; mean IgG of responders 816 mg/dl vs. 475 mg/dl of non-responders). Further, a trend was observed that prior cGvHD impairs vaccination response as 88.9% of the non-responders but only 54.7% of the responders had prior cGvHD (p = 0.073). The results demonstrate, that the currently proposed vaccination strategy leads to seroprotection in the majority of alloHSCT patients.

Brief report: attitudes towards Covid-19 vaccination among hospital employees in a tertiary care university hospital in Germany in December 2020.

Coronavirus disease 2019 (Covid-19) vaccination is essential to fight the pandemic. Health care workers (HCWs) are prioritized to get vaccinated, yet uptake of recommended vaccinations is known to be low in this group. In a tertiary care university hospital with a high number of Covid-19 patients in intensive care, 59.5% of surveyed staff (N = 2454) were willing to get vaccinated, 21.4% were unsure and 18.7% refused. Vaccine hesitancy was higher in female, younger and healthy employees without contact to Covid-19 patients; nurses (53.3%) were much less willing to get vaccinated compared to physicians (82.7%).

Use of Pneumococcal and influenza vaccine in patients with COPD, asthma bronchiale and interstitial lung diseases in south east Germany.

INTRODUCTION: The aim of this study was to analyse use of and adherence to influenza and pneumococcal vaccination in high-risk patients with chronic pulmonary disease. METHODS: The study was initiated at the Centre of Pneumology in Donaustauf, Germany. All patients with asthma bronchiale (AB), chronic obstructive pulmonary disease (COPD) and interstitial lung disease (ILD) that were treated in a pneumological Non-ICU ward, in the sleep laboratory or in the outpatient's clinic between October 1st, 2019 and March 26th, 2020 and provided informed consent were included. Vaccination certificates and a vaccination-centred questionnaire were analysed in relation to vaccination status, risk factors, patient characteristics. RESULTS: 133 patients with COPD, 68 patients with AB and 104 patients with ILD were included. PCV13/PPSV23 vaccination only (no sequential vaccination) was performed in less than 10%/33% of all patients. Sequential vaccination of PCV13 and PPSV23 was performed in 12.8% of COPD, 7.4% of AB patients and 13.5% of ILD patients. Influenza vaccination was performed in less than 30% of all patients. Vaccinations were mainly performed by general practitioners (GPs) and rarely by specialists of pulmonary care (<6%). 67% of all patients were seen by a specialist in pulmonary care in the last 36 months, but in less than 15% the vaccination status was evaluated. DISCUSSION: Use of and adherence for PPSV23 and influenza vaccinations is low in patients with COPD, AB and ILD in south east Germany.

Impact of Parvovirus B19 Viremia in Liver Transplanted Children on Anemia: A Retrospective Study.

Acute parvovirus B19 (B19V) infection in immunocompromised patients may lead to severe anemia. However, in adult transplant recipients, B19V reactivations without anemia and low-level viremia are common. The impact of B19V in pediatric transplant patients, with high risk of primary infection, is investigated here. In a six-month period, 159 blood samples of 54 pediatric liver transplant recipients were tested for B19V DNA by quantitative real-time PCR. Viremia was correlated with anemia and immunosuppression and compared with rates in adult transplant recipients. B19V DNA was detected in 5/54 patients. Primary B19V infections were observed in four patients prior to and in one patient after transplantation. Rates of viremia were significantly higher in pediatric recipients than in adults. Prolonged virus shedding after primary infection prior to transplantation accounts for most viremic cases. Anemia was significantly more frequent in samples from viremic patients, but remained mild. In 15% of anemic samples, B19V DNA was detected. Therefore, in anemic pediatric transplant recipients, diagnostics for B19V seem reasonable.

Low-level DNAemia of parvovirus B19 (genotypes 1-3) in adult transplant recipients is not associated with anaemia.

BACKGROUND: After acute parvovirus B19 (B19V) infection of immunocompetent individuals, viral genomes persist lifelong in various tissues. In immunocompromized patients, acute B19V infection may be associated with severe anaemia. It is unclear whether reactivation of latent B19V DNA may contribute to persistent viraemia and anaemia in transplant recipients. OBJECTIVE AND STUDY DESIGN: We retrospectively analysed the impact of B19V infection in 371 adult transplant recipients (kidney, liver, heart, bone marrow). The patients' pre-transplantation serostatus was determined. 1431 sera or plasmas obtained in monthly intervals during six months following transplantation were analysed for the presence of B19V DNA by quantitative PCR which allows discrimination between B19V genotypes 1-3. RESULTS: Overall, 82% of the patients were seropositive. B19V DNA (<600-1100 geq/ml) was detected in 4.0% of patients and classified as genotype 1 in 12, genotype 2 in one and genotype 3 in two patients. Whereas 5.5%, 6.7% and 5.7% of liver, heart and bone marrow recipients displayed DNAemia, viral genomes were detected only in 1.4% of kidney recipients. Haemoglobin levels and reticulocyte counts showed no differences between DNAemic and non-DNAemic patients. In a control group of 120 healthy subjects, 78% were seropositive and 2.5% displayed DNAemia. CONCLUSIONS: Prevalence and level of B19V DNAemia in adult transplant recipients was comparable to that observed in healthy individuals, but with a distinct accumulation within the first weeks post-transplantation. The presence of low-level DNAemia in transplant recipients was not associated with anaemia.

Detection of hepatitis E virus (HEV) from porcine livers in Southeastern Germany and high sequence homology to human HEV isolates.

BACKGROUND: Hepatitis E virus (HEV) has been identified as an emerging cause of infectious hepatitis over the last years in developed countries. In contrast to travel associated hepatitis E, zoonotic sources of infection are suspected for autochthonous cases in Europe. OBJECTIVE: Since pigs are known reservoirs of HEV, we tested porcine livers sold as food in Southeastern Germany for the presence of hepatitis E virus RNA. STUDY DESIGN: We purchased 200 porcine liver samples in 81 butcher shops and grocery stores in Regensburg, Germany. Nucleic acid preparations were tested for the presence of HEV RNA by quantitative real-time PCR (RT-qPCR). HEV isolates from positive samples were characterized by partial sequencing of ORF1 and ORF2 regions in the HEV genome and by phylogenetic analysis. RESULTS: Specimens from eight (4%) of 200 purchased pig livers had detectable HEV RNA amounts. Sequence determination and phylogenetic analysis allowed two novel isolates to be classified as HEV genotype 3, subgenotype 3a (swR437) and 3c (swR269), respectively. Both novel swine HEV isolates showed high sequence homology to isolates obtained from patients with acute HEV infection from the same geographic region. CONCLUSIONS: These results support the suggested role of undercooked pig products in food as a source of zoonotic HEV infection for humans. It remains to be clarified if this mechanism of transmission is responsible for the surprisingly high anti-HEV IgG prevalence recently observed in some European countries and the USA.

Low risk of Clostridium difficile infections in hospitalized patients with inflammatory bowel disease in a German tertiary referral center.

INTRODUCTION: Many reports, mainly from the US and Canada but also a recent report from a center in Europe, have documented the increasing impact of Clostridium difficile infections in patients with inflammatory bowel disease (IBD) during the last years. To determine the prevalence of C. difficile infections in hospitalized IBD patients in a tertiary referral center in Germany, we conducted this retrospective analysis. METHODS: Data of all IBD in-patients treated due to an acute flare of their IBD at the Department of Internal Medicine I of the University of Regensburg between January 1, 2001, and June 30, 2008, were analyzed. In patients with a concomitant diagnosis of C. difficile infection, further variables such as IBD-related treatment at the time of infection or outcome were examined. RESULTS: In total, 995 in-patients with IBD were treated in this hospital [638 patients with Crohn's disease (CD), 357 with ulcerative colitis (UC)] during the study period. Of these, 279 patients with CD and 242 patients with UC were admitted with an acute flare and suffering from diarrhea and abdominal pain. Only 10 of those were diagnosed as having a concomitant infection with C. difficile. Six patients were female and the median age was 49 years (range: 15-80). Six patients with C. difficile infections suffered from UC and 4 patients from CD, all with previous colonic involvement. Eight patients used immunosuppressive therapies; only 2 patients were treated with antibiotics before infection. CONCLUSION: In contrast to recent reports from other countries, only a low percentage of hospitalized patients with acute flares of their IBD were identified as having an underlying C. difficile infection in this German tertiary referral center. However, in IBD patients with an acute flare, a concomitant C. difficile infection should be excluded, especially in patients with immunosuppressive treatment and colonic involvement of their disease. Further research is needed to evaluate if regions with different risks of C. difficile infections exist and to find out more about potential reasons for this observation.

False-negative serology in patients with acute parvovirus B19 infection.

BACKGROUND: Acute parvovirus B19 (B19V) infection is characterized by high-level viremia. Antibodies against the capsid proteins VP1 and VP2 may complex with B19V-particles thereby becoming undetectable in diagnostic tests. OBJECTIVES: We intended to obtain data on the frequency of false-negative serology in acute B19V-infection. STUDY DESIGN: 129 plasma or serum samples of healthy blood donors and of patients with suspected B19V-infection were analyzed for B19V-DNA by qPCR and VP1/VP2-specific IgG and IgM by ELISA. Eleven of these samples were derived from four pregnant women with previous contact to B19V-infected individuals. Using acidic conditions virus/antibody-complexes were disrupted and detected by WesternLine and ELISA. RESULTS: 83/118 samples were derived from acutely infected individuals displaying viremia (10(3)-10(12)geq/mL). In 24/83 viremic samples (28.9%) VP1/VP2-specific IgM and IgG were undetectable in ELISA, but could be demonstrated to be complexed with B19V-particles. Each 7/83 (8.4%) was IgM-positive/IgG-negative and IgM-negative/IgG-positive, in 45/83 samples (54.2%) IgG and IgM could be detected. 35 samples did not contain B19V-DNA; five of these were from seronegative persons. Analyzing consecutive sera derived from four pregnant women, B19V-DNA was demonstrated in 10/11 samples, B19V-specific IgG- and IgM-antibodies were detectable in 10/11 and 4/11 samples, respectively. In 2/4 women seroconversion was observed, but IgM was not detected in 50% of the samples. B19V-specific IgG but not IgM was detectable in 2/4 women. CONCLUSION: Acute B19V-infection cannot be diagnosed by exclusive analysis of B19V-specific antibodies. Only the combination of assays for detection of B19V-DNA and antibodies enables correct serodiagnosis.

Long-term persistence of tick-borne encephalitis antibodies in adults 5 years after booster vaccination with Encepur Adults.

Tick-borne encephalitis (TBE) is a potentially serious disease, especially in adults. There is no treatment available for TBE; supportive therapy may help to ease symptoms of the disease. Vaccination is the most effective method of preventing TBE disease and is recommended for those who live, work, or travel in TBE-endemic areas. Regular booster vaccinations are recommended every 3-5 years to maintain protection. Evidence from recent clinical studies suggests that TBE antibodies persist at high levels for longer than the current recommended intervals for TBE booster vaccination. The aim of this study was to evaluate the long-term persistence of TBE antibodies in adults after primary vaccination using a rapid schedule and a first booster dose of Encepur Adults, an inactivated TBE vaccine. A total of 222 adults 19-51 years of age were invited for serological follow-up investigations 3 and 5 years following their first booster dose. High antibody titres were recorded throughout the follow-up period. Neutralization test (NT) titres > or =10 were noted in 99% of subjects 3 and 5 years after the first booster vaccination and 97% tested positive by enzyme-linked immunosorbent assay (ELISA). These results indicate that initially high levels of TBE antibodies following the first booster dose of the vaccine may lead to long-term persistence of TBE antibodies, confirming previous findings and suggesting it may be appropriate to extend the interval between booster doses from 3 to 5 years.

Phylogenetic and case-control study on hepatitis E virus infection in Germany.

BACKGROUND: Hepatitis E is a classic water-borne disease in developing countries. In Germany, hepatitis E virus (HEV) infections are notifiable. The number of non-travel-associated infections has increased in recent years, but the route of transmission in most is unknown. Our objective was to determine risk factors for autochthonous HEV infections in Germany. METHODS: Cases of HEV met clinical definitions and were confirmed by laboratory analysis (defined as detection of HEV by polymerase chain reaction [PCR] or immunoglobulin M by serologic testing). PCR products from blood or stool samples were genotyped for phylogenetic analysis. A case-control study included case subjects with autochthonous HEV infection and matched control subjects who were randomly recruited from a population-based telephone list. RESULTS: From May 2006 through August 2007, 76 of 96 persons for whom HEV infection had been reported to the routine surveillance system were interviewed. Sixty-six persons had disease that fulfilled the inclusion criteria: 45 (68%) had autochthonous infection, and 21 (32%) had travel-associated disease. Genotypes 3 or 4 were present in 15 of 15 persons with autochthonous infection, and genotype 1 was present in 8 of 9 persons with travel-associated infection. In conditional logistic regression involving 45 case subjects and 135 control subjects, consumption of offal (41% vs. 19%; odds ratio [OR], 2.7; 95% confidence interval [CI], 1.2-6.2) and wild-boar meat (20% vs. 7%; OR, 4.3; 95% CI, 1.2-15.9) were independently associated with autochthonous HEV infection. CONCLUSION: Hepatitis E is endemic in Germany and likely exists as a food-borne zoonosis. Implicated meat products should be investigated to provide recommendations for preventive measures.

Association of parvovirus B19 infection and Hashimoto's thyroiditis in children.

Hashimoto's thyroiditis is a common autoimmune disorder of the thyroid gland. It has been linked to infections with hepatitis C, EBV, HTLV-1, and Yersinia enterocolitica. As parvovirus B19 has been associated with a wide spectrum of autoimmune diseases, we investigated the potential role of B19 infection in inducing Hashimoto's thyroiditis. Serum samples derived from 73 children and adolescents with Hashimoto's thyroiditis and from 73 age-matched controls were included in the study. The mean age of disease manifestation was 10 y 7 mo. All samples were analyzed for the presence of viral DNA and for antibodies against VP1, VP2, and NS1 proteins. VP1- and VP2-specific antibodies were present in 38 patients (52%) and 43 controls (59%; N.S.). NS1-specific antibodies were detectable in 23 patients (32%) and 19 controls (26%; N.S.). Parvovirus B19 DNA was detectable in 9 patients (12%) and 2 controls (3%; p < 0.03), indicating recent B19-infection. A negative correlation between disease duration and the detection of viral DNA was seen. The mean disease duration in B19-DNA-positive patients was 6 mo, compared to 29 mo in the remainder (p < 0.01). There is strong evidence that acute parvovirus B19 infections are involved in the pathogenesis of some cases of Hashimoto's thyroiditis.