All-in-One Solutions for Detection of Antinuclear Antibodies
When the immune system starts fighting its own body, the autoimmune disease
Lupus erythematodes may be the culprit. It is one of the common autoimmune
diseases that can affect an organ, an organ system or, ultimately, the whole body.
It is difficult to say exactly how many autoimmune diseases exist. Some diseases
are suspected of having autoimmune causes, but it has not yet been possible to
supply proof. The current list contains well over 60 diseases.
The most common autoimmune diseases
• Rheumatoid arthritis
• Lupus erythematodes
• Diabetes mellitus type I
• Hashimoto thyroiditis
• Morbus Graves
• Coeliac disease
• Morbus Crohn
• Colitis ulcerosa
• Multiple sclerosis
• Guillian-Barré syndrome
• Scleroderma
• Sjögren’s syndrome
• Goodpasture’s syndrome
• Morbus Addison
• Wegner’s granulomatosis
• Primary biliary sclerosis
• Sclerosing cholangitis
• Autoimmune hepatitis
• Polymyalgia rheumatica
• Raynaud’s disease
Antinuclear antibodies indicate autoimmune diseases
- Hep-2 D1933 H+: HEp-2 cells positive to Anti Centromere Antibodies
As many autoimmune diseases are difficult to identify,laboratory tests are helpful for confirming the clinical diagnosis. The most commonly used assay is the determination of antinuclear antibodies (ANAs) by indirect immunofluorescence analysis (IFA). ANAs indicate the possible presence of autoimmunity and help the pathologist to specify the diagnosis and initiate appropriate therapy. ANAs can also be found in patients with conditions that are not considered classic autoimmune diseases, such as chronic infections and cancer.
Detection of ANAs requires special slides with sections of specific mouse organs (i.e. stomach, liver, kidney or other, depending on the type of disease to be detected) or slides coated with special cellular lines (Hep-2 or HeLa). The serum of the patient reacts with this substrate, and by adding an anti-human secondary antibody that conjugates with fluoresceinisothiocyanate (FITC) this reaction is visible via fluorescence microscopy. Moreover, antibodies can be detected by a direct reaction between the serum and the pathogenic agent (e.g. virus, bacteria, etc). In this case the slides are coated with specific inactivating substrates for each pathogenic agent.
- Hep-2 D1933 H+: HEp-2 cells positive to antinuclear antibodies – homogeneous pattern
All-in-one solution for immunofluorescence
For IFA analysis all reagents can be purchased as ready-to-use kits from companies that supply diagnostic products for pathology labs. Together with
two leading Italian vendors – Dasit S.p.A. and Bouty S.p.A. (Technogenetics) – Leica Microsystems is going a step further to offer users a genuine all-in-one
solution. The two companies have put together an application package containing all the necessary components for IFA: a microscope with fluorescence axis, a camera, documentation and archiving software – from Leica Microsystems – and reagents.
Positive user feedback
Dasit and Bouty Technogenetics have been extremely successful with this all-in-one solution. However, the most important result of the cooperation is the positive feedback of the customers, who are particularly impressed by the quality of the microscope images and the easy and ergonomic handling of IFA. Even laboratory staff with little experience of microscopic analysis are able to use the system correctly within a surprisingly short time, as the system solution not only has an extremely userfriendly design, but fits seamlessly into the laboratory
workflow.
Comments from Dasit and Bouty Managers
“After various installations we are really satisfied and enthusiastic about the application support offered by Leica Microsystems. The support received
during the internal training was very efficient and has been highly appreciated by our Application Specialists and Sales Representatives.”
“For correct interpretation of ANA positive detections, it is very important to characterise and identify the right “pattern” of the fluorescence distribution, because each pattern corresponds to a different type of disease. With the Leica Microsystems solution we optimised the performance of our IFA reagents, improved the definition of the fluoroscopic pattern and took a big step forward in facilitating clinical diagnosis.”
Economic Microscope System Solution for Immunofluorescence
To provide optimised instrumentation for Immunofluorescence Diagnostics, Leica Microsystems offers a cost effective microscope system solution that consists of a Leica Digital Microscope Leica DM1000 or DM2000 with fluorescence axis and FITC filter cube. An optional LED illumination for fluorescence is also available.
In addition the system can be equipped with a Leica DFC420 digital colour camera for fast and convenient image acquisition. Leica IM or Leica Application Suite (LAS) offer efficient tools for ensuring reliable documentation and image analysis as well as a powerful database for storing every single diagnosis and printing medical reports.