Elisa Assays, imunofluorescenční zařízení a PCR testy
Fotooxidací indukovaný systém zesílení
Peter25 května, 20210 Comments
Fotooxidací indukovaný systém zesílení fluorescence pro ultracitlivý enzymový imunosorbentní test (ELISA)
This report suggests a method of enhancing the sensitivity of chemifluorescence-based ELISA, using photooxidation-induced fluorescence amplification (PIFA). The PIFA utilized autocatalytic photooxidation of the chemifluorescent substrate, 10-acetyl 3,7-dihydroxyphenoxazine (ADHP, Amplex Red) to amplify the fluorescent product resorufin, initially oxidized by horse radish peroxidase (HRP). As the amplification rate is proportional to the initial level of resorufin, the level of antigen labeled by HRP is quantified by analyzing the profile of fluorescence intensity.
The normalized profile was interpolated into an autocatalysis model, and the rate of increase at half-maximum time was quantified by the use of an amplification index (AI). The lower limit of detection, for resorufin or HRP, was less than one-tenth that of the plate reader. It requires only slight modification of the fluorescence reader and is fully compatible with conventional or commercial ELISA. When it is applied to a commercial ELISA kit for the detection of amyloid beta, it is verified that the PIFA assay enhanced the detection sensitivity by more than a factor of 10 and was compatible with a conventional 96-well ELISA assay kit. We anticipate this PIFA assay to be used in research for the detection of low levels of proteins and for the early diagnosis of various diseases with rare protein biomarkers, at ultra-low (pg/mL) concentrations.
Description: A sandwich quantitative ELISA assay kit for detection of Mouse Adiponectin (ADP) in samples from serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids.
Description: A sandwich quantitative ELISA assay kit for detection of Mouse Adiponectin (ADP) in samples from serum, plasma, tissue homogenates, cell lysates, cell culture supernates or other biological fluids.
Description: Quantitative sandwich ELISA kit for measuring Mouse adiponectin, ADP in samples from serum, plasma, cell culture supernates, tissue homogenates, cell lysates. A new trial version of the kit, which allows you to test the kit in your application at a reasonable price.
Description: Quantitative sandwich ELISA kit for measuring Mouse adiponectin, ADP in samples from serum, plasma, cell culture supernates, tissue homogenates, cell lysates. Now available in a cost efficient pack of 5 plates of 96 wells each, conveniently packed along with the other reagents in 5 separate kits.
Description: A competitive ELISA for quantitative measurement of Mouse adiponectin,ADP in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A competitive ELISA for quantitative measurement of Mouse adiponectin,ADP in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: A competitive ELISA for quantitative measurement of Mouse adiponectin,ADP in samples from blood, plasma, serum, cell culture supernatant and other biological fluids. This is a high quality ELISA kit developped for optimal performance with samples from the particular species.
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Mouse Adiponectin (ADP) in serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids.
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Mouse Adiponectin (ADP) in serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids.
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Mouse Adiponectin (ADP) in serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids.
Description: This is Double-antibody Sandwich Enzyme-linked immunosorbent assay for detection of Mouse Adiponectin (ADP) in serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids.
Description: Enzyme-linked immunosorbent assay based on the Double-antibody Sandwich method for detection of Mouse Adiponectin (ADP) in samples from Serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids with no significant corss-reactivity with analogues from other species.
Prevalence strongyloidiázy v kohortě migrantů v Itálii a přesnost nového testu ELISA pro S. stercoralis Infekce, průřezová studie
Strongyloides stercoralis infection is a life-threatening neglected tropical disease. Diagnostic issues have caused an underestimation of its global burden. The choice of appropriate diagnostic tests for the screening of populations at risk of the infection, such as migrants from endemic countries, is of paramount importance. From November 2017 to July 2018, all migrants presenting to the National Institute for Health Migration and Poverty (INMP) in Rome, Italy were offered screening tests for S. stercoralis infection. The study objective was to estimate the prevalence of strongyloidiasis in the study population and the accuracy of a novel ELISA assay.
The following tests were carried out at the IRCCS Sacro Cuore Don Calabria hospital in Negrar, Verona: stool microscopy, real-time PCR for S. stercoralis, in-house immunofluorescence test (IFAT), a commercial ELISA assay (Bordier ELISA), and a novel ELISA assay (Euroimmun ELISA). A latent class analysis (LCA) model set up with test results, clinical variables, and eosinophilia indicated a prevalence around 7.5%, in line with previous findings. The sensitivity and the specificity of Euroimmun ELISA were 90.6% (95% CI 80.5-100) and 87.7% (95CI 84.5-91.0); these results indicate that the novel ELISA assay would be suitable for screening of migrants from endemic countries.
Analýza funkčních virů generovaných PAMP RNA pomocí testu ELISA IFNα / β
Virus-generated PAMP RNAs are key factors that activate host immune response. The PAMP RNAs are therefore usually closely related with viral disease pathogenesis. Quantitative real time PCR is a conventional method to assess RNA. However, it cannot be used for detecting short dsRNAs generated by viral replicase. This protocol was established to analyze the PAMP RNAs produced by viruses which are able to induce host immune response. Classical viral PAMP RNAs and non-classical viral PAMP RNAs are analyzed separately.
Briefly, to access total viral PAMP RNAs, total RNA was extracted from the virus infected cells and then transfected into Cop5 cells. Whereas, to assess non-classical viral PAMP RNAs, the constructs expressing viral replicase are transfected into Cop5 cells. The amount of IFNα/β produced by Cop5 cells, determined by ELISA, is correlated with the total and non-classical viral PAMP RNAs. Since this method is based on type I IFN response, it is therefore suitable for measuring the functional virus-generated PAMP RNAs and also for assessing the efficiency of these PAMP RNAs.
Vývoj sendviče s dvojitou protilátkou a enzymovým imunosorbentem (DAS-ELISA) pro detekci KHV
Koi herpesvirus disease (KHVD) caused by the koi herpesvirus (KHV) is difficult to diagnose in live fish, presenting a challenge to the koi industry. The enzyme-linked immunosorbent assay (ELISA) method cannot be widely used to detect KHV because few commercial anti-KHV antibody exists. Here, we developed an anti-ORF132 polyclonal antibody and confirmed its reactivity via indirect immunofluorescence assay and Western blotting. A double-antibody sandwich ELISA (DAS-ELISA) was established to detect KHV, monoclonal antibody 1B71B4 against ORF92 was used as the capture antibody, and the detection antibody was the polyclonal antibody against the truncated ORF132.
The lowest limit was 1.56 ng/ml KHV. Furthermore, the DAS-ELISA reacted with KHV isolates, while no cross-reactions occurred with carp oedema virus, spring viraemia of carp virus, frog virus 3 and grass carp reovirus. Two hundred koi serum samples from Guangdong, China, were used in the DAS-ELISA test, and the positive rate of the koi sera was 13%. The clinical sensitivity and specificity of the DAS-ELISA relative to the traditional PCR method were 66.7% and 97.6%, respectively. Our findings may be useful for diagnosing and preventing KHVD in koi and common carp.
Enzymově vázaný imunosorbentní test (ELISA) s použitím rekombinantního Fasciola katepsinu L1 pro diagnostiku lidské fasciolózy způsobené hybridním typem Fasciola hepatica / gigantica
Recombinant Fasciola cathepsin L-1 (rCatL1) was evaluated in enzyme-linked immunosorbent assay (ELISA) for the serodiagnosis of human fasciolosis in Japan. Quality characteristics of the test were accessed by receiver operating characteristic (ROC) analysis, with sera from fasciolosis patients (n = 10), patients with no evidence of parasitic infections (n = 29), and patients with other helminth infections (n = 119). Both the sensitivity and specificity of the test achieved 100% with the control samples. To test the performance of the assay in an authentic situation, 311 serum samples, which had been sent to our laboratory for the diagnosis of parasitic infections from January 2018 to February 2019, were re-assessed using the rCatL1 ELISA. In this case, the sensitivity of the rCatL1 ELISA was 100%, giving positive results to all fasciolosis sera (n = 7), and the specificity was 99.0%, in which three of the 304 non-fasciolosis samples were judged positive.
Careful re-examination of the laboratory data and medical imaging of these three patients revealed that one of the patients, who had been diagnosed as having larva migrans syndrome, was judged to be infected with Fasciola, in addition to ascarid nematodes. Thus the true specificity of the assay in the authentic reached 99.3% (302/304). As the rCatL1 ELISA exhibited a highly significant positive likelihood ratio (152.0) and negative likelihood ratio (0.0), calculated from the 311 sample data, this rCatL1 ELISA can be used for routine screening and definitive diagnosis test for fasciolosis in reference laboratories.
Description: AMP-Fluorescein conjugate calibrator is desigend to use with our green fluorescent FAM-cAMP PDE IV substrate (#Cat: 13602), a cAMP derivative that is a specific substrate for phosphodiesterase (PDE) IV.
Description: GMP-Fluorescein conjugate calibrator is desigend to use with our green fluorescent FAM-cGMP PDE V substrate (#Cat: 13604), a cGMP derivative that is a specific substrate for phosphodiesterase (PDE) V.
Description: These antibodies bind to two spike proteins (S1 & S2) on the SARS-CoV-2 virus and can detect presence of SARS-CoV-2 ELISA test COVID-19 spike protein using antibody pair ab-131-295-PAIR. Antibodies: Rabbit anti-spike protein S1 was coated at a dilution of 1/1000 in PBS for 1 hour at room temperature. After blocking, calibrator was diluted from 2000ng/ml to 15ng/m1 in blocking buffer and incubated for 1 hour on coated plate. After washing detection antibody (Chicken anti-spike protein S2 – HRP) was diluted 1/1000 in blocking buffer and incubated on the calibrator wells for 1 hour. After a final wash and tamping, the ELISA was developed by the addition of 100u1TMB to each well and stopped after 15 minutes by the addition of 0.2M sulfuric acid.