PreDicta chip-based high resolution diagnosis of rhinovirus-induced wheeze.

Rhinovirus (RV) infections are major triggers of acute exacerbations of severe respiratory diseases such as pre-school wheeze, asthma and chronic obstructive pulmonary disease (COPD). The occurrence of numerous RV types is a major challenge for the identification of the culprit virus types and for the improvement of virus type-specific treatment strategies. Here, we develop a chip containing 130 different micro-arrayed RV proteins and peptides and demonstrate in a cohort of 120 pre-school children, most of whom had been hospitalized due to acute wheeze, that it is possible to determine the culprit RV species with a minute blood sample by serology. Importantly, we identify RV-A and RV-C species as giving rise to most severe respiratory symptoms. Thus, we have generated a chip for the serological identification of RV-induced respiratory illness which should be useful for the rational development of preventive and therapeutic strategies targeting the most important RV types.

Correction: HIV microarray for the mapping and characterization of HIV-specific antibody responses.

Correction for 'HIV microarray for the mapping and characterization of HIV-specific antibody responses' by Daniela Gallerano et al., Lab Chip, 2015, 15, 1574-1589.

HIV microarray for the mapping and characterization of HIV-specific antibody responses.

We used the microarray technology to develop chips containing a comprehensive set of proteins and peptides covering the proteome of HIV-1 clade C, which is the HIV-1 subtype that causes the majority of infections worldwide. We demonstrate that the HIV microarray allows simultaneous, sensitive and specific detection of antibody responses for the major immunoglobulin classes (IgG, IgA, IgM, IgE) and subclasses (IgG1-4) with minute amounts of serum samples towards a large number of HIV antigens and peptides. Furthermore, we show that the HIV chip can be used for the monitoring of antibody responses during the course of the disease and during treatment. The HIV microarray should be useful to study antibody responses to multiple HIV antigens and epitopes in HIV-infected patients to explore pathomechanisms of the disease, for diagnosis and for monitoring of treatment and of vaccine trials.

Advances in allergen-microarray technology for diagnosis and monitoring of allergy: the MeDALL allergen-chip.

Allergy diagnosis based on purified allergen molecules provides detailed information regarding the individual sensitization profile of allergic patients, allows monitoring of the development of allergic disease and of the effect of therapies on the immune response to individual allergen molecules. Allergen microarrays contain a large variety of allergen molecules and thus allow the simultaneous detection of allergic patients' antibody reactivity profiles towards each of the allergen molecules with only minute amounts of serum. In this article we summarize recent progress in the field of allergen microarray technology and introduce the MeDALL allergen-chip which has been developed for the specific and sensitive monitoring of IgE and IgG reactivity profiles towards more than 170 allergen molecules in sera collected in European birth cohorts. MeDALL is a European research program in which allergen microarray technology is used for the monitoring of the development of allergic disease in childhood, to draw a geographic map of the recognition of clinically relevant allergens in different populations and to establish reactivity profiles which are associated with and predict certain disease manifestations. We describe technical advances of the MeDALL allergen-chip regarding specificity, sensitivity and its ability to deliver test results which are close to in vivo reactivity. In addition, the usefulness and numerous advantages of allergen microarrays for allergy research, refined allergy diagnosis, monitoring of disease, of the effects of therapies, for improving the prescription of specific immunotherapy and for prevention are discussed.

Exercise with latex sport bands represents a risk for latex allergic patients.

Based on two clinical observations of adverse reactions during exercise with latex sport bands, we aimed to assess the possible risk for allergic patients posed by this equipment by investigating allergen content and IgE binding potential. Protein extracts of three different latex sport bands were characterized with sera of latex allergic patients. The IgE recognition profile of the allergic patients was identified by component resolved diagnosis and the allergen composition of the extracts was characterized by inhibition assays with the recombinant latex allergens Hev b 1, 3, 5, 6.02, and 8. The sera showed pronounced IgE binding to all three blotted extracts, however with diverse patterns. Inhibition assays revealed the presence of Hev b 1, 3, 5, and 8 in latex sport band extracts. The clinical relevance of contained allergens was demonstrated by strong skin reactions when testing with latex sport bands. From our results we conclude that latex sport bands contain clinically relevant allergens and may cause latex allergic individuals to experience allergic symptoms, potentially amplified by exercise-induced mechanisms. Even though latex is labeled on products, it is important that patients as well as athletic trainers and physical therapists recognize the risk of adverse reactions with these bands.

Frequency of 23 human papillomavirus types using DNA microarray in women with and without cytological anomalies.

OBJECTIVE: To evaluate the frequency of single and multiple human papillomavirus (HPV) infections in women with and without cervical dysplasia. MATERIALS AND METHODS: An oligonucleotide microarray system was used to detect 19 types of high-risk HPV (HPV-16/-18/-311-33/-35/-39/-45/-51/ -52/-53/-56/-58/-59/-66/-68/-73/-821-16 variant E-E6-G350/-16 variant E-E6-T350) and 4 types of low-risk HPV (HPV-6/-11/ -42/-44) in 122 consecutive women visiting our colposcopy outpatient clinic classified into controls (normal epithelium, nonspecific cervicitis, metaplasia; n = 56) and cervical intraepithelial neoplasia (CIN) (n = 66). RESULTS: In 78/122 (64%) cervical samples, HPV DNA was detected. Compared to controls, HPV infection was significantly more prevalent among women with CIN (8/56 [14%] versus 49/66 [74%]; p = 0.001). HPV-18 and HPV-16 were the most common HPV types in all specimens (25% [31/122] and 25% [31/122], respectively). Of note, HPV-16 was significantly more frequent in women with CIN compared to controls (35% [23/66] vs. 14% [8/56], respectively; p = 0.02). Double HPV infections were detected in 16/122 (13%) and multiple infections in 43/122 (35%) women. Multiple HPV infections were found significantly more often among women with CIN compared to controls (30/66 [45%] vs. 13/56 [23%], respectively; p = 0.002). Using a univariate and multivariate logistic regression model to estimate the relative risk of CIN vs. HPV type, HPV-16-positive cases were found to have the highest risk of CIN (odds ratio [OR] 3.2; 95% confidence interval [CI] 1.3-7.9; p = 0.002). CONCLUSION: Multiple HPV infections are common in women with and without CIN, but significantly more prevalent among women with CIN compared to controls.

Allergen microarrays for the diagnosis of specific IgE against components of cow's milk and hen's egg in a multiplex biochip-based immunoassay.

Over the last few decades, the prevalence of allergic diseases has increased dramatically in developed nations. The resulting burden on health care systems worldwide has provoked a whole series of research initiatives among allergy experts and commercial companies that aim to develop novel tests to improve the diagnostic risk assessment and early preventive treatment of the disease. The advent of protein microarray technology has inspired the development of miniaturized immunological applications that permit the simultaneous analysis of huge numbers of disease-related parameters. Allergen microarrays have been developed for the monitoring of patient-specific antibody profiles to a previously unknown variety of allergens in a single analytical step. This has been accomplished by the successful adaptation of solid-phase antibody assays for the detection of surface-bound allergens to the microarray format, the development of appropriate assay conditions, and the improvement of software-guided microarray image analysis. Here we report a protocol for the development and analysis of food allergen microarrays.

Protein microarrays for the diagnosis of allergic diseases: state-of-the-art and future development.

In the emerging field of Functional Proteomics, protein microarrays are considered to be one of the most promising tools for the simultaneous analysis of the a) abundance, b) function, and c) interaction of proteins on a system-wide scale. Resting on the technological grounds of widely used DNA biochips, the great power of microarray-based miniature solid-phase immunoassays lies in their potential to investigate in parallel large numbers of analyte pairs in a variety of biological samples. Consequently, this has fueled aspirations that protein microarrays may serve as tools for the high-throughput functional investigation of complete proteomes and, moreover, that they will develop into promising candidates for innovative in-vitro diagnostic (IVD) applications. To date, published examples of protein microarrays for IVD purposes have included tests for allergy, autoimmune and infectious diseases. Here, we discuss recent advancements in the development of protein microarrays for the profiling of IgE antibodies in the diagnosis of Type 1-related allergic diseases.

Microarray-based detection of mannose-binding lectin 2 (MBL2) polymorphisms in a routine clinical setting.

The assessment of allelic variants in the human mannose-binding lectin 2 (MBL2) gene is of great clinical importance in newborns or immune-suppressed patients at high risk for a variety of infections. Here, we present a study on the genotyping accuracy of a DNA microarray-based on-chip PCR method suited for the detection of five different polymorphisms in the MBL2 gene. We tested 153 genomic DNA samples, prepared from archival blood spots on Guthrie cards, for the presence of allelic variants in the human MBL2 gene by the on-chip PCR method and compared the obtained results of three variants to standard DNA capillary sequencing. The genotyping power of the described assay was readily comparable to DNA sequencing (453/459 correct genotype calls in 153 DNA samples; 98.7% accuracy), mainly due to intrinsic technical benefits of microarrays such as high number of test replicates and automated data analysis. This study demonstrates, for the first time, the accuracy and reliability of a microarray-based on-chip PCR genotyping assay for measuring allelic variants in a routine clinical setting.

Affinity determinations of purified IgE and IgG antibodies against the major pollen allergens Phl p 5a and Bet v 1a: discrepancy between IgE and IgG binding strength.

Allergen-specific IgE and IgG antibodies coexist in allergic individuals, but only IgE has anaphylactogenic capacity. This study aimed to determine the association, dissociation and equilibrium constants for the interaction of allergen-specific IgE and IgG with the major grass and birch pollen allergens Phl p 5a and Bet v 1a. We isolated specific IgE and IgG antibodies from pollen allergic patients' sera by a two-step affinity chromatography protocol and controlled the high purity in a recombinant allergen chip microarray. Surface plasmon resonance measurements of polyclonal IgE and IgG species revealed that their affinities diverge widely, being in the range of 10(-10) and 10(-11) M for IgE, but only 10(-6)-10(-7) M for IgG. Moreover, murine monoclonal IgG1 antibodies against the allergens showed affinities of 10(-7)-10(-8) M. Thus, we conclude from our data that even stringently affinity matured IgG cannot score the superior affinity of IgE antibodies to allergens.

Protein microarrays in diagnosing IgE-mediated diseases: spotting allergy at the molecular level.

Over the last few decades, the prevalence of allergic diseases has increased dramatically in developed nations. The resulting worldwide burden on healthcare systems has provoked a whole series of research initiatives among allergy experts and commercial companies that aim to develop novel tests to improve the diagnostic risk assessment and early preventive treatment of disease. The advent of protein microarray technology has fueled aspirations of multianalyte immunological applications that permit the simultaneous analysis of huge numbers of disease-related parameters that will hopefully become amenable in the near future. Allergen microarrays have been developed for the monitoring of patient-specific antibody profiles to a previously unknown variety of allergens in a single analytical step. This review describes significant discoveries and developments in allergy research against a background of the increasing prevalence of disease and hence the emerging challenges for national healthcare systems. The development of novel protein microarray-based allergy diagnostic tests is portrayed in concert with the recent advances and benefits of this technology, along with the challenges that must be met by manufacturers in order to succeed with innovative allergen microarrays in a highly competitive market.

Microarrayed allergens for IgE profiling.

Diagnosis of type I allergy is based on anamnesis, provocation testing, and serological determination of total and specific IgE. Currently, in vivo and in vitro diagnostic tests employ allergen extracts prepared from various allergen sources (e.g., pollen, mites, animal dander, moulds, foods, venoms, etc.). The application of recombinant DNA technology to the field of allergen characterization has allowed to reveal the molecular nature of the most common allergens. To date a continuously increasing number of allergen sequences has become available and panels of recombinant allergens-assembling the epitope complexity of natural allergens sources-can be produced. The use of recombinant allergens instead of crude, natural extracts for allergy diagnosis allows us to determine the individual IgE reactivity profile of each patient. To enable a comprehensive analysis of the patient's IgE binding pattern to a large number of individual allergens, a new type of serological test is required. In this paper, we applied microarray technology to create a multi-allergen test system, based on microarrayed recombinant allergens.

Microarrayed allergen molecules: diagnostic gatekeepers for allergy treatment.

Type I allergy is an immunoglobulin E (IgE)-mediated hypersensitivity disease affecting more than 25% of the population. Currently, diagnosis of allergy is performed by provocation testing and IgE serology using allergen extracts. This process defines allergen-containing sources but cannot identify the disease-eliciting allergenic molecules. We have applied microarray technology to develop a miniaturized allergy test containing 94 purified allergen molecules that represent the most common allergen sources. The allergen microarray allows the determination and monitoring of allergic patients' IgE reactivity profiles to large numbers of disease-causing allergens by using single measurements and minute amounts of serum. This method may change established practice in allergy diagnosis, prevention, and therapy. In addition, microarrayed antigens may be applied to the diagnosis of autoimmune and infectious diseases.