Designing a novel ELISA method based on CagA, NapA recombinant antigens to increase sensitivity and specificity of Helicobacter pylori whole cell antigen detection

Aim: In this research, we designed a direct Enzyme Linked Immunoassay method to detect Helicobacter pylori antigens in stool specimens

Background: Helicobacter pylori infection as the worldwide problem is related to many gastrointestinal disorders such as gastritis, gastric cancer, non-ulcer disease, peptic ulcer disease and duodenal ulcer

Methods: We produced and purified recombinant CagA and NapA antigens in Escherichia coli and extracted their antibodies from a panel of positive sera specimens. We designed a novel enzyme linked immunoassay direct method in combination with the whole cell for the qualitative and quantitative detection of Helicobacter pylori antigens in human stool. Assay performance was evaluated by histopathology staining and urease activity

Results: The sensitivity and specificity of assay was determined as 91.7 [95% confidence interval: 89.3-95.6%] and 93.1% [95% CI: 91.2-96.4%], respectively. Novel ELISA exhibits enhanced sensitivity and specificity of Helicobacter pylori detection in comparison with another commercially available kit

Conclusion: Combination of the recombinant antigens and whole cell of Helicobacter pylori in immunoassay designing is a new approach about early diagnosis, treatment and fallowing up of the Helicobacter pylori infected patients, especially in peptic cancer cases

spectrum of diseases caused by aspergillus fumigates

The opportunistic mould Aspergillus is a ubiquitous fungus. Diseases caused by Aspergillus species are most commonly caused by Aspergillus fumigatus. The spectrum of disease caused by Aspergillus is dependent on the health of the immune system. The ranges of illnesses individuals acquire are aspergilloma, allergic bronchopulmonary aspergillosis, invasive aspergillosis, sinusitis, otomycosis, ocular infections, CNS infection, osteomyelitis, cutaneous aspergillosis, endocarditis, urinary tract infection. Aspergilloma is the most common clinical presentations of lung infections due to Aspergillus species. Allergic Bronchopulmonary Aspergillosis is a result of an immune reaction to colonization of Aspergillus fumigatus within the airways of patients. Invasive aspergillosis is generally seen in severely immunocompromised individuals. Aspergillus sinonasal infections may or may not be invasive and can follow a fulminant or an indolent course. Otomycosis has typically been described as fungal infection of the external auditory canal. Aspergillus endophthalmitis may occur by several mechanisms, including direct inoculation by trauma after surgical procedures or by hematogenous spread. Central nervous system [CNS] aspergillosis is a rare and uniformly fatal complication of disseminated disease, involving the cerebral hemispheres and cerebellum. The mechanism for Aspergillus bone infections is by direct extension, traumatic injury, inoculation by a surgical intervention, hematogenous spread and injection drug abusers. Primary cutaneous disease is a rare disease caused by Aspergillus fumigatus. Aspergillus species have been reported as a cause of both native and prosthetic valve endocarditis. Aspergillosis of urinary tract may occur by three ways namely, by ascending infection from the lower tract, from haematogenous dissemination or due to Aspergillus cast in renal pelvis. Keywords: Aspergillus fumigatus, disease, aspergilloma, invasive aspergillosis

[Production of camel polyclonal antibody against vascular endothelial growth factor receptor-2 and performance evaluation in ELISA test]

In molecular approach, serum of camel contains a unique type of antibodies devoid of light chains since the light chain is missing, the heavy-chain antibodies should bind their antigen by one single domain, the variable domain of the heavy immunoglobulin chain. Vascular endothelial growth factor receptor-2 [VEGFR-2] is one of the important proteins in angiogenesis which over-expressed in many types of tumors. Two male dromedaries [Camelus dromedarius] were immunized against VEGFR-2. ELISA was used to evaluate the immunization process. Camel immune sera were fractionated with protein A and G affinity chromatography. Heavy chain antibodies tested for its specific reactivity in cell-based ELISA in recognition of VEGFR-2 on the cell surface of three cell lines; 293/KDR, HUVECs and A431. In ELISA test, the camel sera in 1/12800 dilution had the acceptable results. Furthermore, cell-based ELISA demonstrated that the polyclonal heavy chain antibodies bind to VEGFR-2 in 293/KDR and HUVECs cell lines. Single chain polyclonal antibodies against VEGFR-2 can be used for detection of soluble form of this protein by ELISA, and also flowcytometry, western blotting and immunohistochemistry

Effects of gamma irradiation on proliferation and IL-5 production of peripheral blood lymphocytes

Gamma irradiation is routinely used for suppression of lymphocyte function in transfusion and transplantation procedures. In recent years, some investigators focused on the effects of ionizing radiation on special aspects of lymphocyte function and considered the possibility of its clinical application for treatment of some immunological disorders. In this study, we evaluated the effects of five different doses of