Testing
When it comes to the detection of Lyme and coinfections, the main type of test is a blood test which looks for antibodies to the bacteria that cause the infectous disease. This is known as Serology. If antibodies have not developed sufficiently or the immune system is not strong enough to develop antibodies (surpressed immunesystem), it is possible for these tests to be negative despite active infection, which is referred to as a “false negative”. Tests have always been a controverse part of the detection process and guilty of many sufferes not having being given the adequate treatment due to false negatives or incomplete detection of the pathogen and infection.
Types of tests
The are two basic tuses of tests, indirect or direct. The difference is where the focus is, on immune response or on infection.
Indirect: Measures immune response to the infection (Serologies and T-cell response assays).
Direct: Detects and measures the infection organism itself.
tests
FISH (Fluorescent in-situ hybridization)
Fluorescence in situ hybridization (FISH) is a technique used in genetic testing to help diagnose diseases caused by chromosomal differences. Healthcare providers can also use FISH to detect gene changes that they can target to treat the pathogen. FISH uses proteins or chemical labels to “paint” parts of chromosomes or genes that pathologists look for to diagnose diseases. The labels look like colored lights under a microscope.
Elisa
Enzyme-linked Immunoabsorbent Assay. This test system can be both indirect and direct.
Direct ELISAs use a conjugated primary antibody, while indirect ELISAs include an additional amplification step. In an indirect ELISA, an unconjugated primary antibody binds to the antigen, then a labeled secondary antibody directed against the host species of the primary antibody binds to the primary antibody.
Western Blot
The western blot (sometimes called the protein immunoblot), or western blotting, is a widely used analytical technique in molecular biology and immunogenetics to detect specific proteins in a sample of tissue homogenate or extract. Besides detecting the proteins, this technique is also utilized to visualize, distinguish, and quantify the different proteins in a complicated protein combination. Indirect.
ImmunoBlot
The ImmunoBlot, also known as Western blot, is an advanced method for identifying proteins in biological samples. In the case of infections, this method is often used to confirm the presence of specific antibodies against certain proteins or antigens. First, the proteins are separated by electrophoresis according to their size and then transferred onto a solid support, such as a nitrocellulose membrane. The sample is then applied to the support, and if specific antibodies are present, they bind to the corresponding proteins. Subsequently, the bound antibodies are visualized, often by adding an enzyme- or fluorescence-labeled secondary antibody and an appropriate substrate. The band patterns generated by the immunoblot can provide information about the presence and specificity of antibodies in the sample.
EliSpot
EliSpot is a technology used to assess the functional immune response of T cells to various antigens through enzyme-linked immune spot assays. This test system enables the examination of a variety of infections and pathogens, including well-known agents such as Ehrlichia & Anaplasma, Bartonella, Babesia, as well as viral infections caused by Epstein-Barr virus (EBV), herpes simplex virus, cytomegalovirus, and others.
iSpot
The iSpot test analyzes T-cell reactions to provide insights into the cellular immune system, crucial for diagnosing chronic and opportunistic infections. It measures cytokines IFN-γ and IL2, and is relevant for various infectious diseases including Borrelia, Anaplasma, and SARS-CoV-2. It aims to set a new diagnostic standard.
TickPlex
The TickPlex assay is conducted based on an ELISA. Compared to other ELISA assays, this test includes a new antigen for Borrelia persisters. Testing with the TickPlex Plus detects antibodies against the pathogens Ehrlichia, Mycoplasma, Bartonella, Rickettsia, Babesia, EBV, Parvovirus, and Coxsackie viruses using the more sensitive and specific ELISA compared to all previously available IFT testing methods. The TickPlex can be used in various stages of disease.
PCR
Polymerase chain reaction (PCR) detects small amounts of genetic material of a pathogen (disease-causing organism) or abnormal cells in a sample of your blood, saliva, mucus, or tissue. The genetic material could be DNA, which contains the genetic information needed for a person and most other living things to develop and grow.
ddPCR
A new direct detection method, the multiplex ddPCR assay, will improve the efficiency and accuracy of direct testing for bartonellosis, babesiosis, and Lyme borreliosis, allowing healthcare providers and patients to be more confident in diagnoses. Instead of performing a single reaction on a test sample, this powerful technology partitions the sample into approximately 20,000 droplets and analyzes each droplet for the presence of DNA. By reducing the potential number of PCR inhibitors in each reaction, the sensitivity of direct detection for low-abundance pathogens is greatly increased. This test method is still not commercially available but will soon be.
Test chart
Relationship between indirect and direct tests:
1.- If immune system is weak, serologies may be false negative, but a weak immune system may allow an increase in the number of organisms, so the direct tests like FISH and PCR may be more sensitive.
2.- Reverse is also true – PCR and FISH negative may be sero-positive due to high immune response.
Since no test is 100% sensitive, what is recommended is a panel approach where testing of both indirect and direct is perfomed.
| Test | Type | Analisis |
| Elisa | Indirect | Serology and T-cell |
| Western Blot | Indirect | Serology and T-cell |
| ImmunoBlot | Indirect | Serology and T-cell |
| EliSpot | Indirect | Serology and T-cell |
| iSpot | Indirect | Serology and T-cell |
| TickPlex (Elisa method) | Indirect | Serology and T-cell |
| FISH | Direct | Infection detection |
| PCR | Direct | Infection detection |
| ddPCR | Direct | Infection detection |
The new generation
| Test | Laboratory | Web |
| ImmunoBlot | IGeneX | https://igenex.com/ |
| FISH | IGeneX | https://igenex.com/ |
| ddPCR | Galaxy | https://www.galaxydx.com/ |
| EliSpot, iSpot, ImmunoBlot, Tickplex, Elisa | Armin Labs | https://arminlabs.com/en/ |
