The MELISA® test - MELISA Medica Foundation

BIOHELLENICA June 2012
The MELISA® test
A revolutionary blood test used to improve diagnosis and
treatment of chronic disease
A licensing opportunity for
Biohellenica
The MELISA® test provides important information for patients suffering
from chronic degenerative diseases, including chronic fatigue syndrome
(CFS), fibromyalgia and multiple sclerosis (MS). It is virtually impossible to
avoid exposure to environmental pollutants, such as metals like nickel,
mercury and titanium. The most common sources of exposure are dental
materials, orthopedic implants and food additives.
With increasing awareness about the possible side-effects from metal
exposure in sensitive individuals comes a growing demand for the
MELISA® test across the world. In Greece we work with several clinics that
send patients for MELISA® testing abroad, but the time and cost for
transport limits the demand.
Metal allergy plays an
important role in a variety of
medical specialties, such as:
• Orthopedics and
implants
• Dentistry
• Occupational medicine
and environmental
health
• Diseases of
immunologic origin
• Infectious diseases
All licensed MELISA® laboratories receive access to our contact network
and comprehensive marketing support; from ready-made marketing
materials to constant exposure through our website www.melisa.org and
through lectures and international conferences.
“Our case demonstrated the need for proper investigation with tests such
as MELISA® and also the need for proper history in all such patients.”
Orthopaedic surgery in a patient with metal sensitivity.
Adala R et al. Aesthet Surg. 2011 Jan-Apr; 4(1): 67-68.

BIOHELLENICA June 2012
The effect of metals on health
The adverse effect metals may have on human
health is a growing area of research and a
connection between metals and the onset or
worsening of symptoms has been documented
by both clinics and in peer reviewed literature.
Metal allergy is usually diagnosed by patch
testing, but this can be problematic when
testing irritative substances as mercury, or
substances with large particles like titanium
dioxide. Using MELISA® avoids many of
obstacles of patch testing.
Chronic exposure to metals in jewellery, dental
implants and restorations, cosmetics or joint
prostheses can sensitize genetically
predisposed individuals and induce a so-called
Type IV allergic reaction (cellular type
hypersensitivity).
Metals are potential allergens. The binding of
metal ions to proteins can alter the protein
activity and surface structure of these
enzymes, thereby inducing local or systemic
symptoms characteristic of allergic reactions or
autoimmune diseases.
In contrast to a Type I allergy, which is
mediated by IgE antibodies, a Type IV allergic
reaction is mediated by T lymphocytes that
have had prior contact to a given allergen
(memory lymphocytes). Following renewed
contact with the sensitizing allergen, these
memory lymphocytes respond by enlarging
(lymphoblast transformation) and dividing
(proliferation). The newly formed cells (effector
cells) together with their secreted cytokines
mediate the resulting inflammatory reaction.
It is important to differentiate between
metal allergy and metal toxicity. The
MELISA® test does not measure the levels of
metals in a patient’s body; it measures whether
the patient is allergic to metals. For example,
hair samples may show levels of mercury or
other substances which are below the official
“safe limit” – but the patient can still be
allergic. For allergic individuals, there is no such
thing as a “safe” limit. Even a minute amount of
a substance may pose danger in an allergic
individual.
A well-known allergen, nickel is present
in some dental materials and nickelcontaining
alloys. Such exposure can
cause serious health problems in nickelallergic
patients. Nickel is also present in
cheap jewelry and many household
items. Nickel-allergic patients may
benefit from a low nickel diet, mainly
avoiding foods such as nuts, beans and
oats.
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BIOHELLENICA June 2012
Orthopedic surgery and implants
Metals are found in body implants such as hip joints,
knee prosthesis, screws, pacemakers, silicone breast
implants, cochlear implants and dental implants. In the
majority of patients, these implants are biocompatible.
However, in sensitive patients, the implants may
induce side-effects such as pain, dermatitis, cutaneous
swelling, impaired wound healing, bone infections and
implant loosening. Less knows is that even excessive
fatigue can be due to chronic inflammation due to
metal ions.
Although bacterial infection is often the cause of
implant rejection, sometimes no bacteria are detected
despite of inflammation. These adverse effects are
mediated by so called wearing products of implant
materials. Nickel, cobalt and chromium are known
metal allergens.
Recently, the cases of titanium allergy have also been
described in the literature. Titanium is a transition
metal and thus may function as a hapten and trigger
cellular hypersensitivity. Since titanium is used as white
pigment in toothpaste, cosmetics and medicaments,
the sensitization of susceptible individuals is possible.
Dentistry
Mucosal changes around dental metal fillings such as
oral lichen may be due to an allergic reaction to the
metal component. Replacement of fillings in metalsensitive
patients leads to disappearance of the
symptoms (Prochazkova et al. 2004).
Allergic reactions to metals used in dental prosthesis,
implants (for example titanium implants, see picture
above right) and braces can be measured in the
MELISA® test. Symptoms may not always be local but
can be systemic and include inflammation in different
body parts.
Occupational medicine and
environmental health
The MELISA® test can be used to screen workers who
are exposed to metals, chemicals or other allergens at
work. If sensitivity is detected, the worker should be
placed in an allergen-free environment to avoid the
development of sometimes serious diseases.
In the case of berylliosis, a sarcoidosis-like disease, a
beryllium-specific blood test is used as a golden
standard by US industry to screen workers for allergy
against beryllium. It has been found that asymptomatic
workers with memory cells to beryllium will - if
beryllium exposure continues - develop a serious lung
disease. This will not happen if asymptomatic
beryllium-sensitized workers are relocated to a
beryllium-free environment. In addition to lifelong
suffering, considerable savings in sick leave costs are
obtained. Certain occupations pose a special risk for
inducing metal allergy. These include construction
workers, miners, electricians, rubber/wood/paper/
textile industry workers, dentists, hairdressers and
painters.
Patients with known clinical metal sensitivity such as
adverse reactions to gold and nickel containing
earrings should consider non-metallic restorative
materials.
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BIOHELLENICA June 2012
Diseases of immunologic origin
Metal allergy is common in patients with various diseases such as skin
diseases (psoriasis, eczema), autoimmune diseases (Multiple Sclerosis,
thyroiditis, Sjögren’s disease) and gastrointestinal diseases. Patients with
symptoms of profound fatigue of unclear etiology (Chronic Fatigue
Syndrome, Myalgic Encephalitis) often suffer from metal allergy induced
by dental metals. Candida allergy, drug allergy (such as penicillin allergy)
and allergy to chemicals (such as formaldehyde) can also be tested. Metal
allergy has also been implicated in autistic disorders and allergy against
thimerosal (a mercury preservative in vaccines) can be detected by the
MELISA® test.
Avoidance of allergens often results in significant health improvement of
the patient. Follow-up MELISA® testing usually shows reduced
lymphocyte reactivity as well.
Diagnosis of active Lyme Borreliosis
A recent development of the MELISA® test involves diagnosis of Lyme
disease / active Borreliosis. Standard laboratory testing includes the
serological test ELISA, which has very low specificity, and the Western
blot, which is very specific. These tests can give false negative results in
patients with early disease and therefore a misdiagnosis can occur.
MELISA® detects cellular immuno reactivity of active infections of Borrelia
burgdorferi . The test improves laboratory diagnosis by confirming an
active infection in patients with or without clinical symptoms.
Further applications
MELISA® can also test for allergy to gluten, gliadin and casein, and testing
to other food substances is being developed.
MELISA® test
procedure
Blood in citrate tubes must
arrive to the laboratory
optimally within 24 hours after
the blood has been drawn,
maximally within 48 hours.
White blood cells (lymphocytes)
are isolated and tested against
allergens chosen accordingly to
the patient’s anamnesis and
exposure to metals. A
questionnaire, which goes
through the most common
sources of metal exposure, is
available for download at
www.melisa.org. The blood is
incubated for five days and the
lymphocyte reaction is measured
by ways: one based on the
uptake of radioisotope by
dividing lymphocytes; the other
by evaluation by microscopy.
The level of reactivity is
measured as a Stimulation Index
(SI). A value over 3 indicates a
positive reaction to a given
allergen. The results are
available in ten days.
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BIOHELLENICA June 2012
Licensing of the MELISA® test
Certified laboratories may perform the MELISA® test under a license agreement. The
MELISA® test is protected by trademark. Currently, laboratories performing the test are
found Australia, Belgium, Germany, South Africa, Spain, Switzerland, United Kingdom
and USA.
Laboratory equipment and material needed for
MELISA® testing
Cytocentrifuge
CO 2 incubator
Table centrifuge for spinning of 50 ml falcon tubes
Betacounter for counting of radioactivity on filters
Harvester for DNA
Different multipipettes
Sterile bench (Laminoflow)
Microscope for light microscopy incl. objective 25x and
40x
Invert microscope (optional)
Water bath
The testing requires disposable material such as cell
media, human serum, radioactive thymidine, plastic
ware (pipettes, tubes, plates), metals solutions etc.
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BIOHELLENICA June 2012
Case study
Sue Collins was 10 years old when her first amalgam fillings
were placed. She suffered from urticaria and rashes
throughout her teenage years and suspected nickel allergy.
As an adult, she underwent tubal ligation with a metal clip.
She developed gallstones and had gall bladder removed.
After placement of dental metal crowns, she experienced
intense abdominal pains and was diagnosed with Irritable
Bowel Syndrome. Additional crowns and bridgework were
fitted and she experienced increasing muscle pain and
weakness. Finally she received the diagnosis of
fibromyalgia and chronic fatigue syndrome. The MELISA ®
test showed a positive reaction to several metals in found
in her dental restoration and metallic clips. After having the
metals removed, her oral symptoms, IBS and fibromyalgia
disappeared.
“I can honestly say I feel like a
new person;
I have my life back.“
Reference literature
Some articles are available from www.melisa.org/articles.php
Hypersensitivity reactions to metallic implants - diagnostic
algorithm and suggested patch test series for clinical use.
Schalock PC et al, Contact Dermatitis. 2012 Jan;66(1):4-19.
Orthopaedic surgery in a patient with metal sensitivity.
Adala R, Chakravarthy M, Srinivas V, Pai S. J Cutan Aesthet
Surg. 2011 Jan-Apr; 4(1): 67-68.
Metal allergens of growing significance: epidemiology,
immunotoxicology, strategies for testing and prevention.
Forte G, Petrucci F, Bocca B. Inflamm Allergy Drug Targets.
2008 Sep;7(3):145-62.
The role of environmental factors in autoimmune
thyroiditis.
Hybenova M, Hrda P, Procházková J, Stejskal V,
Sterzl I. Neuro Endocrinol Lett. 2010;31(3):283-9.
LTT-MELISA ® is clinically relevant for detecting and
monitoring metal sensitivity. Valentine-Thon E, Müller K,
Guzzi G, Kreisel S, Ohnsorge P, Sandkamp M. Neuro
Endocrinol Lett 2006; 27(Suppl 1): 17-24
Hypersensitivity to titanium: Clinical and laboratory evidence.
Müller K, Valentine-Thon E. Neuro Endocrinol Lett 2006;
27(Suppl 1): 31-35
Validity of MELISA ® for metal sensitivity testing. Valentine-
Thon E., et. al, Neuroendocrinology Letters,
2003;24(1/2):57–64
A novel lymphocyte transformation test (LTT-MELISA ) for
®
Lyme borreliosis. Valentine-Thon E et al., Diagn Microbiol Infect
Dis. 2006 Jul 27
MELISA Diagnostics Limited
Diagnosis and treatment of metal-induced side-effects. Stejskal
V, Hudecek R, Stejskal J, Sterzl I. Neuro Endocrinol Lett 2006;
27(Suppl 1): 7-16
The role of metals in autoimmunity. Stejskal J, Stejskal V.
Neuroendocrinology Letters 1999; 20:351-364
Metal-specific lymphocytes: biomarkers of sensitivity in man.
Stejskal, V, et al. Neuroendocrinology Letters 1999; 20:289-
298
Removal of dental amalgam and other metal alloys
supported by antioxidant therapy alleviates symptoms and
improves quality of life in patients with amalgam-associated
ill health. Lindh U, Hudecek R, Danersund A, Eriksson S,
Lindvall A. Neuro Endocrinol Lett. 2002;23(5-6):459-82.
Nickel allergy is found in a majority of women with chronic
fatigue syndrome and muscle pain - and may be triggered by
cigarette smoke and dietary nickel intake. Regland B,
Zachrisson O, Stejskal V, Gottfries CG. Journal of Chronic
Fatigue Syndrome, Vol. 8(1) 2001
Mercury-Specific Lymphocytes: An indication of Mercury
Allergy in Man. Stejskal V, Forsbeck M et al, Journal of
Clinical Immunology, Vol. 16, No 1, 1996
MELISA ® - an in vitro tool for the study of metal allergy.
Stejskal V, Cederbrant K, Lindvall A, Forsbeck M, Toxicology
In Vitro, 1994, vol 8, p. 991-1000
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