Jörg Berg

Kepler University Hospital / Kepler Universitätsklinikum Linz · Med. Campus 3. Institut für med.-chem. Labordiagnostik / Med. Campus 3. Institute of Laboratory Medicine

Background: Genotyping for clinically important single nucleotide polymorphisms (SNPs) is performed by many clinical routine laboratories. To support testing, quality controls and reference materials are needed. Those may be derived from residual patient samples, left over samples of external quality assurance schemes, plasmid DNA or DNA from cell...

The widespread use of molecular PCR-based assays in analytical and clinical laboratories brings about the need for test-specific, stable, and reliable external controls (EC) as well as standards and internal amplification controls (IC), in order to arrive at consistent test results. In addition, there is also a growing need to produce and provide s...

Background Determination of the hepatitis C virus (HCV) genotype and discrimination between HCV subtypes 1a and 1b is still mandatory prior to anti-HCV treatment initiation. The aim of this study was to evaluate the performance of the recently introduced cobas Methods The cobas Results When accuracy was tested, panel members containing HCV subtyp...

A 47-year old women the patient exhibited atypical chest pain. She had a history of resuscitation due to torsade de pointes tachycardia caused by a long QT-syndrome. Therefore, a single chamber ICD had been implanted. There were no clinical signs of systemic infection such as fever or abnormal fatigue. Echocardiography showed a very large walnut sh...

Background: The purpose of this study was to investigate the applicability of the Greiner Saliva Collection System (SCS) to obtain human genomic DNA for the analysis of single nucleotide polymorphisms (SNP) in the clinical routine laboratory. Methods: Saliva and EDTA-blood were collected pair-wise from 112 participants. DNA was prepared by two a...

Acute myeloid leukemia (AML) comprises a spectrum of myeloid malignancies which are often associated with distinct chromosomal abnormalities, and the analysis of such abnormalities provides us with important information for disease classification, treatment selection and prognosis. Some chromosomal abnormalities albeit recurrent are rare such as te...

We developed a sequencing assay for genotypic HIV-1 tropism determination. The assay allows examination of HIV RNA from plasma and HIV DNA from peripheral blood mononuclear cells (PBMC), including PBMC samples from patients with undetectable viral loads. Assessment of 100 pairs of plasma and PBMC samples showed a high concordance of 90%. With the l...

We developed a sequencing assay for genotypic HIV-1 tropism determination. The assay allows examination of HIV RNA from plasma and HIV DNA from peripheral blood mononuclear cells (PBMC), including PBMC samples from patients with undetectable viral loads. Assessment of 100 pairs of plasma and PBMC samples showed a high concordance of 90%. With the l...

To investigate relapse predictors in chronic hepatitis C (CHC) patients with end-of-treatment response (ETR), after pegylated interferon-alpha (PegIFN-alpha) and ribavirin treatment. In a retrospective study we evaluated a spectrum of predictors of relapse after PegIFN-alpha and ribavirin treatment in 86 CHC patients with ETR. Viral loads were dete...

B-type natriuretic peptide (BNP) plays a key role in the regulation of volume homeostasis, and elevated blood levels of BNP are associated with end-stage renal disease. Renal transplantation leads to a decrease of elevated BNP levels with established graft function. Assessment of N-terminal pro-BNP (NT-proBNP) is established as reflecting volume ho...

Vitiligo is a disorder of pigmentation associated with an autoimmune-mediated loss of melanocytes from the epidermis. Humoral immunity and the involvement of cellular immunity have been investigated in the pathogenesis of vitiligo. We evaluated the role of pro-inflammatory cytokines and lymphocyte fractions in peripheral blood in a cohort of Austri...

A sequencing assay for detection of mutations conferring resistance to human immunodeficiency virus type 1 (HIV-1) integrase inhibitors raltegravir and elvitegravir was developed using the automated TruGene sequencing system. The assay returned clear sequencing results for samples with ≥500 RNA copies/ml for mutation detection and HIV-1 subtyping a...

Pulse wave velocity (PWV) is at least partially controlled by vascular tone. Vascular tone and underlying physiological processes such as sympathetic activity, plasma catecholamin, and cortisol levels have been shown to follow diurnal variations. Carotid-to-radial PWV was non-invasively assessed by applanation tonometry in 21 young (26.5+/-2.3 year...

Arterial stiffness is thought to play a critical role in the pathogenesis of cardiovascular events, and in hyperthyroidism increased cardiovascular event rates have been reported. To investigate markers of systemic arterial stiffness, volume homeostasis, and subendocardial perfusion and its interrelationship in patients with Graves' disease (GD) in...

Prevalence of insulin resistance (IR) is increased in type 2 diabetes and in end-stage renal disease (ESRD). IR is associated with advanced atherosclerosis and is an independent predictor for cardiovascular disease in diabetes and ESRD patients. We investigated prevalence, severity, predictors and relation to vascular diseases by the homeostasis mo...

Hypolactasia and lactose intolerance are common conditions worldwide. Hypolactasia seems to be strongly correlated with genotype C/C of the genetic variant C-->T(-13910) upstream of the lactase phlorizin hydrolase (LPH) gene. We developed a rapid genotyping assay for LPH C-->T(-13910) and investigated the relationship of positive lactose breath hyd...

Pertussis, also called whooping cough, is caused by Bordetella pertussis . The disease may show an atypical course, particularly in neonates and elderly patients. A rapid and safe diagnostic method is thus essential for appropriate treatment and prophylaxis. Culture has been considered the gold standard for detection of B. pertussis , but this meth...

To establish and evaluate a new test system for rapid detection and diagnosis of adenoviral keratoconjunctivitis. After establishment of the molecular assay, 52 conjunctival smears were studied. Samples were derived from patients with a clinical presentation compatible with keratoconjunctivitis. A molecular assay for detection of human adenovirus (...

A 56 year old man presented with ichthyosis vulgaris since early childhood, clinically characterised by fine scaling of the trunk and hyperkeratotic scales on the exterior surfaces of the upper and lower extremities. The patient also showed hypothyroidism due to hypoplastic thyroid, cataract, hypercholesterinemia with concommitant arcus cornealis a...

Arterial stiffness is at least partially controlled by vascular tone. Vascular tone and underlying physiological processes, e.g. sympathetic activity, have been shown to follow diurnal variations. This study investigated whether arterial stiffness and perfusion of subendocardial myocardium relative to cardiac workload show diurnal variations under...

A molecular assay for parallel detection of three bacteria, Chlamydia (C.) pneumoniae, Legionella (L.) spp., and Mycoplasma (M.) pneumoniae, in clinical specimens by a set of real-time polymerase chain reactions (PCRs) in a single run was evaluated. Bacterial DNAs were extracted by an automated DNA extraction protocol on the MagNA Pure LC System. A...

Real-time PCR assays are widely used for the detection and quantification of pathogen-derived nucleic acids in clinical samples (1)(2)(3). Because clinical specimens contain PCR-inhibitory moieties that are not always reliably removed during nucleic acid extraction, real-time PCR assays are furnished with internal amplification controls (IACs) to i...

Atherosclerosis is looked upon as an inflammatory disease. The production of proinflammatory markers may indicate activity in this inflammatory state. We prospectively evaluated a range of proinflammatory serum parameters in 136 cardiac patients who had previously undergone percutaneous coronary intervention (PCI). By means of myocardial scintigrap...

The surface topography of red blood cells (RBCs) was investigated under near-physiological conditions using atomic force microscopy (AFM). An immobilization pro- tocol was established where RBCs are coupled via molecular bonds of the membrane glycoproteins to wheat germ agglutinin (WGA), which is covalently and flexibly tethered to the support. Thi...

Little is known about the pathogenic role and the endemic situation of transfusion transmitted virus (TTV). In this study, a molecular assay for detection of TTV based on automated nucleic acid extraction and real-time PCR was developed and evaluated. The new assay includes an internal control. After optimization of the molecular assay, 103 clinica...

Reverse transcription (RT)-PCR is a very sensitive tool for the detection and quantification of RNA including mRNA and is widely used in research and in diagnostic applications (1, 3). RT-PCR assays include RNA controls that are co-analyzed in parallel or within the same reaction tube as internal controls or as quantification standards (1). As an a...

Herpes viruses represent important causes of morbidity and mortality especially in immuno-compromised patients. To assist in rapid diagnosis real-time PCR assays have been developed for the detection of herpes virus DNA in patient specimens. A recently described set of real-time PCR assays using LightCycler technology enabled parallel detection of...

We report on the development of a fully automated real-time PCR assay for the quantitative detection of hepatitis B virus (HBV) DNA in plasma with EDTA (EDTA plasma). The MagNA Pure LC instrument was used for automated DNA purification and automated preparation of PCR mixtures. Real-time PCR was performed on the LightCycler instrument. An internal...

Molecular detection of herpes simplex virus (HSV) DNA is recognized as reference standard assay for the sensitive and specific diagnosis of central nervous system and genital infections caused by HSV. In this study, a qualitative molecular assay based on automated DNA extraction on the MagNA Pure LC (Roche Applied Science, Mannheim, Germany) and a...

Molecular assays based on real-time PCR are widely used for the detection and quantification of virus-derived nucleic acids in clinical specimens (1). When quantification is performed with real-time PCR, sample nucleic acids are commonly tested against reference nucleic acids used as external quantification standards. The external quantification st...

Cardiovascular morbidity and mortality is markedly increased in patients with end-stage renal disease (ESRD) undergoing hemodialysis (HD) and is further pronounced when diabetes mellitus is also present. As atherogenesis is mediated by inflammation of vessel walls and as evidence evolves that atherosclerosis and diabetes mellitus share a common inf...

We read with interest the favorable findings of Holzl et al. (1) showing good agreement of plasma HIV-1 RNA measurements (by the ultrasensitive COBAS AMPLICOR HIV-1 Monitor test) by using the conventional (manual) RNA purification method and the fully automated method performed with the MagNA Pure LC instrument. Their findings would enable diagnost...

Genotyping of hepatitis C virus (HCV) is clinically relevant to epidemiology, prognosis, and therapeutical management of HCV infection. Accuracy and specificity of three assays for HCV genotyping/subtyping were determined. The TruGene HCV 5'NC Genotyping Kit (TruGene), which is a direct sequencing test and two assays based on reversed hybridization...

Autoimmune thyroid diseases are thought to be mediated by pro-inflammatory cytokines such as TNFalpha and IL-6. Serum levels of cytokines may indicate activity levels of immune functions. We investigated serum levels of IL-6 and of the soluble receptor of TNFalpha in patients with newly diagnosed onset of Graves' hyperthyroidism. The predominantly...

The ultrasensitive COBAS AMPLICOR HIV-1 Monitor test was complemented with automated RNA purification on the MagNA Pure LC instrument. This enabled entirely automated ultrasensitive assessment of viral loads in human immunodeficiency virus type 1 (HIV-1)-infected individuals. The detection limit of the fully automated assay and the viral load measu...

Real-time polymerase chain reaction (PCR) assays allow convenient detection and quantitation of virus-derived nucleic acids in clinical specimens. When specimens are assayed for the presence of virus-derived nucleic acids against external standards, sample adequacy is not monitored. This can be achieved by using internal controls that are co-amplif...

Human herpes viruses cause a spectrum of diseases that are usually self-limiting but can be reactive during immuno-suppression and may then lead to severe or even life-threatening diseases. The LightCycler technology allows rapid polymerase chain reaction (PCR) including product analysis within a closed system. This approach has been demonstrated t...

The real-time PCR technology allows convenient detection and quantification of virus derived DNA. This approach is used in many PCR based assays in clinical laboratories. Detection and quantification of virus derived DNA is usually performed against external controls or external standards. Thus, adequacy within a clinical sample is not monitored fo...

The development of a novel normalized quantitative competitive real-time PCR on the LightCycler instrument (NQC-LC-PCR) and its application to the quantification of cytomegalovirus (CMV) DNA in clinical samples are described. A heterologous competitor DNA was spiked into test samples and served as an internal amplification control. The internal con...

Molecular assays for qualitative detection of Legionella spp. in clinical specimens were evaluated. DNA extraction was done either with a fully automated DNA extraction protocol on the MagNA Pure LC System or with manual DNA extraction. Amplification and detection were done by real-time polymerase chain reaction (PCR) on the LightCycler (LC) instru...

For the detection of hepatitis C virus (HCV) specific nucleic acids the polymerase chain reaction (PCR) is widely used. Rapid-cycle PCR is performed in glass capillaries with the LightCycler instrument and allows PCR including product analysis to be performed within a closed system in about 1 h. Thus, rapid-cycle PCR appears especially suitable for...

As serotonin is a mediator of inflammatory joint disease, serum levels were investigated in human patients with arthritis for a possible corresponding role as a disease marker. 48 patients were evaluated by bone scan for disease activity. 5-HT and CRP were measured in the whole group, and IL-6 in those not yet receiving corticosteroids. The pro-inf...

We present the clinical case of a 20-year-old male soldier who appeared in general good physical condition. He suffered from infectious mononucleosis caused by Epstein-Barr virus that had recurred 2 years after the first serologically documented episode. The detected splenomegaly persisted in the healthy young man, who otherwise showed no apparent...

For many diagnostic applications, the specificity and sensitivity of polymerase chain reaction (PCR) is markedly enhanced by applying two rounds of PCR with nested or semi-nested pairs of primers. In two-round PCR protocols on the LightCycler instrument, amplification products must be collected from the capillaries by centrifugation, a procedure th...

To investigate whether the genes encoding Cyclooxygenase-1 and -2 are expressed in thyroid epithelial cells, in vitro. COX-1/-2 gene expression was examined in the thyroid epithelial cell line Nthy-ori3-1 using semi-quantitative RT-PCR and Western blot analysis. ELISAs were employed to assess whole cell COX-enzyme activity, PGE2 and IL-6 formation....

HN-56249 (3-(2,4-dichlorothiophenoxy)-4-methylsulfonylamino-benzenesulfonamide), a highly selective cyclooxygenase (COX)-2 inhibitor, is the prototype of a novel series of COX inhibitors comprising bicyclic arylethersulfonamides; of this series HN-56249 is the most potent and selective human COX-2 inhibitor. HN-56249 inhibited platelet aggregation...

To investigate anti-inflammatory effects of lornoxicam in vitro on COX-1/COX-2, on NO formation from iNOS and on the formation of the pro-inflammatory cytokines TNF-alpha, IL-1beta, IL-6, and IL-8. COX-1 inhibition in intact cells was assessed employing two systems: measurement of aggregation in human washed platelets and assessment of TXB2 formati...

NSAIDs inhibit the conversion of arachidonic acid into Prostaglandin G2 and Prostaglandin H2 which is catalyzed by the enzyme cyclooxygenase (COX). Two genetically distinct isoforms have been discovered, COX-1 and COX-2. While COX-1 is thought to account for homeostatic amounts of eicosanoids, COX-2 is induced during inflammation leading to patholo...

The two isoforms of cyclooxygenase (COX-1, COX-2) are considered the rate-limiting enzymes in the pathway of the conversion of arachidonic acid to eicosanoids (1). COX-1 is constitutively expressed whereas COX-2 is highly up-regulated by various stimuli in inflammatory controlling cells leading to increased eicosanoid formation. Increased eicosanoi...

Excess eicosanoid formation during inflammation has been attributed to the expression of the gene coding for the inducible isoform of prostaglandin G/H synthase (PGHS-2). Human and murine PGHS-2 proteins differ in 73 out of the 604 amino acids. When comparing the inhibitory effects of a panel of PGHS-inhibitors in a whole cell human and murine PGHS...

In controlling inflammation, monocytes and macrophages not only secrete various cytokines but also eicosanoids. Since the human monocytic cell line, Mono Mac 6, represents cells at the stage of mature blood monocytes, cells of this line were investigated to find out whether they also resemble mature monocytes with respect to inflammation. In respon...

An HeLa-LAV cell line was established by infecting and subcloning previously described CD4-expressing HeLa cells with HIV-1. Cells of this line stably synthesize all major HIV proteins, release infectious particles of HIV-1, but do not die even after long term culture. More than 90% of the cells express the envelope protein gp120 on the surface. Th...

Although AIDS patients lose human immunodeficiency virus (HIV)-specific cytotoxic T cells, their remaining CD8-positive T lymphocytes maintain cytotoxic function. To exploit this fact we have constructed bispecific antibodies that direct cytotoxic T lymphocytes of any specificity to cells that express gp120 of HIV. These bispecific antibodies compr...

Immunoglobulin genes are generated during differentiation of B lymphocytes by joining gene segments. A mouse pre-B cell contains a functional immunoglobulin heavy-chain gene, but no light-chain gene. Although there is only one heavy-chain locus, there are two light-chain loci: kappa and lambda. It has been reported that kappa loci in the germ-line...

Immunoglobulin genes are generated during differentiation of B lymphocytes by joining gene segments. A mouse pre-B cell contains a functional immunoglobulin heavy-chain gene, but no light-chain gene. Although there is only one heavy-chain locus, there are two lightchain loci: κ and λ.It has been reported that κ loci in the germ-line configuration a...

When termination codons were introduced into exons of the gene for Ig mu chain, steady-state levels of mu mRNA were reduced, both at the pre-B cell stage and at the plasma cell stage. A termination codon in the variable region gene segment and a termination codon in the second exon of the constant region gene segment had effects of similar magnitud...

Cianidanol (Ci) [(+)-catechin] is a lipophilic compound which interacts with membrane lipids and affects responsiveness and function of immunocompetent cells. We therefore studied the immunomodulating properties of Ci on the proliferative response of human peripheral T-cells in one-way mixed lymphocyte reaction (1-MLR) and autologous mixed lymphocy...

The influence of Cianidanol (Ci), a cytoprotective radical scavenger, on peripheral blood mononuclear cells (PBMC) was assessed with respect to its immunomodulatory function. In previous studies performed in our laboratory, a bidirectional influence of Ci on the immune response was observed, depending on its concentration. In order to elucidate thi...