Name of Product

Fasciola hepatica – IgG ELISA

Catalog Number

AF 9650

Short Info

Fasciolosis is one of the most frequently encountered autochthonous helminthic infections in Central Europe. The immunodiagnosis of Fasciola hepatica infections is challenged by high serological cross-reacitivity with other (hepatic) parasitological infections encountered in Central Europe, such as alveolar echinococcosis, toxocarosis and ascariosis, but also other parasitic infections acquired during overseas travel. The SAP-2 recombinant antigen shows a high specificity, especially with patients with other parasitic infections. It is a suitable serological test for routine diagnosis of human fasciolosis, particularly if the results are supported by clinical history.

This product is manufactured by Bordier Affinity Products in Switzerland and distributed in Germany exclusively by Milenia Biotec.

Method/Platform

ELISA in microplate format

Range/Assay Sensivity

77% sensitivity, 98% specificity

Test Principle

The kit provides all the material needed to perform 96 enzyme-linked immunosorbant assays (ELISA) on breakable microtitration wells sensitized with Fasciola hepatica recombinant antigen. Specific antibodies in the sample will bind to the antigen and washing will remove unspecific antibodies. The presence of parasite specific antibodies is detected with a Protein A – alkaline phosphatase conjugate. A second washing step will remove unbound conjugate. Revealing bound antibodies is made by the addition of pNPP substrate which turns yellow in the presence of alkaline phosphatase. Color intensity is proportional to the amount of Fasciola hepatica specific antibodies in the sample. Potassium phosphate is added to stop the reaction. Absorbance at 405 nm is read using an ELISA microplate reader.The test can be performed with automatic systems, but this must be validated by the user.

Brief Instructions

Storage

2-8° C

Components

ELISA wells, dilution buffer, washing solution, control sera, conjugate, substrate solution, stopping solution

12 x 8 strips (96 tests)
Fasciolosis is one of the most frequently encountered autochthonous helminthic infections in Central Europe. The immunodiagnosis of Fasciola hepatica infections is challenged by high serological cross-reacitivity with other (hepatic) parasitological infections encountered in Central Europe, such as alveolar echinococcosis, toxocarosis and ascariosis, but also other parasitic infections acquired during overseas travel. The SAP-2 recombinant antigen shows a high specificity, especially with patients with other parasitic infections. It is a suitable serological test for routine diagnosis of human fasciolosis, particularly if the results are supported by clinical history.

Overview

• Isolate high quality DNA from a broad variety of phage strains
• High yields of total DNA
• Fast and easy processing using a rapid spin-column format
• No phenol or chloroform extractions or cesium chloride banding required
• High yields of DNA recovered 3-15 µg DNA from 10⁶-10¹⁰ pfu/ mL of enriched phages

This kit provides a rapid spin column method for the purification of total DNA from a broad spectrum of bacteriophages propagated in bacteria grown in liquid cultures. The DNA is isolated without the use of phenol, chloroform or cesium chloride banding procedures. The spin-column based procedure is rapid and can be completed in less than 45 minutes. The kit is highly efficient for processing small volumes of phage supernatant (500 µL – 1 mL) and with the optional DNase and Proteinase K treatments phage DNA yields are maximized while host DNA contamination is minimized. Purified total phage DNA is of the highest integrity, and can be used in a number of downstream applications including PCR, qPCR, Restriction Fragment Length Polymorphism (RFLP), sequencing, cloning, Southern Blot and more.

Details

Supporting Data

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* Average yields will vary depending upon a number conditions used and developmental stage.

Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 1 year after the date of shipment.

Introduction

Blood samples contain rich DNA, including mitochondrial DNA, genomic DNA, circulating DNA (mostly released into blood after tumor cell apoptosis) in white blood cells, as well as parasitic viral or microbial DNA. These DNA are important parameters in clinical testing or diagnosis, which are also valuable materials for medical research. There are three main issues with extracting DNA from blood samples:

1. The sample is highly infectious, posing great harm to operators and the environment.

2. The source of DNA is complex and aportion of the nucleic acid, such as viral DNA or free DNA, may be lost during the operation, leading to downstream detection failure;

3. Blood sample contains a large amount of impurities and inhibitory factors.

Currently there are many methods available for extracting DNA from whole blood samples, such as phenol chloroform extraction, salting out method, etc. However, these methods require pre-treatment of blood sample, which removes red blood cells and isolate white blood cells in the first step. Due to the requirement that it cannot inactivate or kill pathogens during the process of removing red blood cells, the waste liquid (red blood cell lysate) and consumables may be contaminated by pathogens and become infectious, posing a danger to the entire laboratory environment and operators. In addition, during the process of removing red blood cells, useful nucleic acid information such as viruses, microorganisms, or circulating DNA is also lost, leading to experiment or detection failures.

The HiPure Blood DNA Kits series provided by Magen Company uses silica gel column purification technology, which can directly lyse whole blood samples without the need for white blood cell separation. Whole blood samples are directly mixed with lysates and proteases, resulting in the inactivation of pathogens, greatly reducing the infectivity, environmental pollution, and the chance of operators being infected. Due to the direct lysis and digestion of samples, except lymphocyte DNA, other circulating DNA as well as DNA from viruses and microorganisms, can also be recovered.

This product provides fast and easy methods for purification of total DNA for reliable PCR and Southern blotting. Total DNA (e.g., genomic, viral, mitochondrial) can be purified from whole blood, tissue and culture cells.

Details

Specifications

Principle

This product is based on silica column purification. The sample is lysed and digested with lysate and protease, DNA is released into the lysate. Transfer to an adsorption column. Nucleic acid is adsorbed on the membrane, while protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, Nucleic acid was finally eluted with low-salt buffer (10mm Tris, pH9.0, 0.5mm EDTA).

Advantages

• High quality DNA – meet a variety of downstream applications, including PCR, qPCR, enzyme digestion, hybridization, etc.
• Fast – without separation of leukocytes, organic extraction or ethanol precipitation
• Simple – all nucleic acids can be obtained by direct digestion
• Pertinence – specially designed for isolating DNA from 3-10ml blood and related body fluids
• Wide applicability – handle a variety of liquid samples
• Economy – excellent cost effectiveness performance

Kit Contents

Storage and Stability

Proteinase K, RNase A should be stored at 2-8°C upon arrival. However, short-term storage (up to 12 weeks) at room temperature (15-25°C) does not affect their performance. The remaining kit components can be stored at room temperature (15-25°C) and are stable for at least 18 months under these conditions.

Purchase Guide

*Note：Leukocyte protocol can be used when large volume whole blood samples need to be processed. Whole blood was treated with red blood cell lysate, and white blood cells were obtained by centrifugation before extraction

Blood samples contain rich DNA, including mitochondrial DNA, genomic DNA, circulating DNA (mostly released into blood after tumor cell apoptosis) in white blood cells, as well as parasitic viral or microbial DNA. These DNA are important parameters in clinical testing or diagnosis, which are also valuable materials for medical research. There are three main issues with extracting DNA from blood samples: