Detection of Anti-Platelet Glycoprotein Antibodies Using MAIPA Method

AITP mostly occur in children accompanied by variable clinical sings including petechiae, purpura, ecchymosis and severe bleeding. This study has determined and characterized the anti-platelet glycoproteins in children with ITP. The aim of this study was to determinate anti-platelet glycoproteins (GPs) using MAIPA method. During 18 months 38 children with clinical signs of AITP were studied in Mofid children hospital. To determine anti-platelet antibodies by ELISA technique, washed O negative platelets were used as a source of platelet antigens. MAIPA method was used to detect antibodies against individual platelet membrane glycoprotein. The anti-platelet antibodies level above mean+ 3SD of control group was assumed as positive. The results indicated that the platelet count ranges was between 2×109/L and 95×109/L. 63.5 % out of 38 patients were anti-platelet antibodies positive with ELISA method. The correlation between the above patients with anti-platelet antibody positive and clinical signs was 0.4. Results for determination of antibody against platelet GPIIb/IIIa, GPIb/IX and GPIa/IIa using MAIPA method were 44%, 51% and 25% respectively. In conclusion the preference of MAIPA method is the detection of very small amount of antibody. Since MAIPA is the specific method for the detection of antibody against glycoprotein antigens, it has the advantage of differentiating immune and non-immune thrombocytopenia

Comparison of recombinant A2-ELISA with rKE16 dipstick and direct agglutination tests for diagnosis of visceral leishmaniasis in dogs in Northwestern Iran

INTRODUCTION: Various methods are used for the diagnosis of visceral leishmaniasis (VL), such as microscopic examination, culture and inoculation of laboratory animals; however, serological assays are commonly used for the detection of antibodies in serum samples with a wide range of specificity and sensitivity. METHODS: The purpose of this study was to compare three serological methods, including rA2-ELISA, the recombinant KE16 (rKE16) dipstick test and the direct agglutination test (DAT), for the detection of antibodies against VL antigens. The assays utilized 350 statistically based random serum samples from domestic dogs with clinical symptoms as well as samples from asymptomatic and healthy dogs from rural and urban areas of the Meshkinshahr district, northwestern Iran. RESULTS: Samples were assessed, and the following positive rates were obtained: 11.5% by rKE16, 26.9% by DAT and 49.8% by ELISA. The sensitivity among symptomatic dogs was 32.4% with rKE16, 100% with DAT and 52.9% with ELISA. Conversely, rA2-ELISA was less specific for asymptomatic dogs, at 46.5%, compared with DAT, at 88.9%. CONCLUSIONS : This study recommends rA2-ELISA as a parallel assay combined with DAT to detect VL infection among dogs. Further evaluations should be performed to develop an inexpensive and reliable serologic test for the detection of Leishmania infantum among infected dogs

Serodiagnosis of recently acquired Toxoplasma gondii infection in pregnant women using enzyme-linked immunosorbent assays with a recombinant dense granule GRA6 protein.

Indirect immunoglobulin G (IgG) and IgM enzyme-linked immunosorbent assays (ELISAs) with a recombinant GRA6 protein of Toxoplasma gondii were developed and evaluated for accurate diagnosis of recently acquired infection in pregnant women. According to the results from Toxoplasma serodiagnostic tests, women were classified into 3 groups representing acute (group I), chronic (group II), or no Toxoplasma infection (group III). To discriminate group I from group II sera, the GRA6-IgG-ELISA reached sensitivity and specificity of 87.5% and 94.1%, respectively. Although 22 (91.7%) of 24 group I sera were positive by the GRA6-IgM-ELISA, only 1 (2.9%) of 34 group II sera scored positive. The GRA6-IgM-ELISA displayed a meaningful correlation with Vidas Toxo IgM and exhibited higher specificity (97.1%) than Euroimmun IgM ELISA (88.2%) (Euroimmun, L?k, Germany) for detection of recent infection. These results demonstrate that IgG and IgM ELISA with rGRA6 are useful to identify and discriminate recent from past Toxoplasma infection in pregnant women

Histopathological changes induced by Hemiscorpius lepturus scorpion venom in mice.

International audienceEnvenomation by Hemiscorpius lepturus (H. lepturus) is associated with local necrosis, followed by systemic manifestations. In this work the LD₅₀ of H. lepturus venom were determined by subcutaneous (SC) injection in white Balb/c mice (5 mg/kg). Histopathological alterations in organs such as kidney, heart, liver, lungs, stomach and intestine were determined in 3, 6, 12 and 24 h following experimental (SC) envenoming injection of one LD ₅₀ of the venom in Balb/c mice. Histological studies showed degenerative changes in the kidney with disorganized glomeruli and necrotic tubular in 3 h and reached to its climax in 6 h. Myocardium showed massive myocytolysis with interstitial necrosis in 3 h and reached to its peak after 6 h past envenoming. Bowels showed edema of lamina propria and slight villous necrosis. The enzymatic activities of creatine kinase (CK) and lactate dehydrogenase (LDH) were significantly increased in the serum in 9 h. No necrotic lesion observed in lungs and liver. The results indicate that the venom of H. lepturus is a highly cytotoxic, and induces massive tissue damages in specific organs, starting from the heart and kidney as the first target in 3 h and ends to the bowels in 6 h post envenomation