Porcine Heavy Chain Immunoglobulin ELISA Kit

In this paper, we report on the demonstration of a portable immunoassay system consisting of a small centrifugal microfluidic device driver (bento box) and a centrifugal microfluidic device made of polypropylene and fabricated by injection molding. The bento box consists of a cheap DC motor and an Arduino microcontroller. It has a simple structure and is the size of a bento box, that is, 150 x 150 x 100 (W x D x H) mm(3). The developed device can automatically execute an enzyme-linked immunosorbent assay (ELISA) process under a steady rotating condition because it was designed based on the principle of CLOCK, which we previously presented. Here, we first executed an ELISA using a system consisting of the bento box and a device made of polydimethylsiloxane (PDMS) and compared it with a servo-controlled device driver. It was confirmed that the results of the bento box were consistent with those of the servo-controlled device driver. The limit of detection (LOD) using the bento box was 0.759 ng ml(-1). Therefore, the controllability of the bento box was demonstrated. Next, we evaluated the injection-molded device through multi-step fluid control. We confirmed, through real-time observation of the device, that accurate flow control in the designed ELISA procedure was executed. Lastly, ELISA was employed for the measurements of mouse IgG using the system consisting of the bento box and the polypropylene device. The system performed all fluidic controls within 12 min; we confirmed the specificity of the system, and the LOD was 0.320 ng ml(-1).

Background. Human interleukin- (IL-) 1 receptor-associated kinase 4 (IRAK-4) deficiency is a recently described primary immunodeficiency. It is a rare, autosomal recessive immunodeficiency that impairs toll/IL-1R immunity, except for the toll-like receptor (TLR) 3- and TLR4-interferon alpha (IFNA)/beta (IFNB) pathways.Case Report. We report the first patient in Greece with IRAK-4 deficiency. From the age of 8 months, she presented with recurrent infections of the upper and lower respiratory tract and skin abscesses. For this, she had been repeatedly hospitalized and treated empirically with intravenous antibiotics. No severe viral, mycobacterial, or fungal infections were noted. Her immunological laboratory evaluation revealed low serum IgA and restored in subsequent measurements; normal IgG, IgM, and IgE; and normal serum IgG subclasses. Peripheral blood immunophenotyping by flow cytometry and dihydrorhodamine (DHR) test revealed normal counts. She was able to make functional antibodies against vaccine antigens, including tetanus and diphtheria. She was administered with empirical IgG substitution for 5 years until the age of 12 years, and she has never experienced invasive bacterial infections so far. DNA analysis revealed a heterozygous variant in the patient: c.823delT (p.S275fs*13 at protein level) in the IRAK4 gene.Conclusions. The importance of clinical suspicion is emphasized in order to confirm the diagnosis by IRAK4 gene sequencing and provide the appropriate treatment for this rare primary immunodeficiency, as soon as possible.