Today's Applications of Immunodiagnostics

Requirements for Future Diagnostic Devices

Nowadays, a lot of diseases can be treated well. A crucial factor fast diagnosis essential for an immediate onset of appropriate treatment. It depicts a critical factor for the patient's health and life and significantly determines his well-being. Moreover, improved diagnostics are not only required regarding the health of a patient: 70% of healthcare expenses are linked to diagnostic tests ¹⁶⁾ . Therefore, improvements in diagnostic technologies have the potential to drastically reduce overall healthcare costs while at the same time increasing health as such.

Diagnostic tests are usually developed for use in air-conditioned laboratories with refrigerated storage of chemicals, a constant supply of calibrators and reagents, highly trained personal and rapid transportation of samples. This setting is not available for most developing countries ²⁾ . Thus, most of the substantial progress achieved in the public health and Point-of-Care sector has only been advantageous to the more developed part of the world.

According to the WHO 2.5 out of 6 billion people lack basic sanitation, 2 billions do not have access to electricity and more than 1 billion lack basic healthcare services and clean drinking water ¹⁹⁾ . Moreover, 50% of all deaths in the most impoverished developing countries are a result of infectious diseases, whereas in the wealthiest developed countries this concerns less than 5% ¹⁾.
Therefore, transforming existing technologies into portable applications is a leap forward to improve general health all over the world. These applications should be robust and sensitive enough for the use outside of equipped laboratories. Outbreaks and spreading of potential epidemic diseases or sexually transmitted infections could be controlled by a rapid diagnosis and immediate onset of appropriate treatment ²⁾.

A need for such technologies is urgent: 500 million people between the age of 15 and 49 are infected with curable sexually transmitted infections like chlamydia, gonorrhea, syphilis or trichomoniasis each year ²⁰⁾ ! However, the infrastructure currently available for the diagnosis of infectious diseases often proves to be too slow and expensive to be practicable for third world countries. This can be illustrated with the identification of pathogens of an infectious diarrhea taking 2-4 days – even in the best developed laboratories of the world ²¹⁾ .

According to the problems and needs discussed, devices for future diagnostics should meet the following requirements:

• Speed - a fast diagnosis reduces time until the beginning of treatment, preventing the spread of epidemic diseases and reducing the severity of a disease


• Simplicity - the handling necessary to perform the test should be as easy as possible


• Low-cost - Point-of-Care diagnostics need to be affordable in developing countries


• Unambiguity - the output of Point-of-Care tests requires the clarity of a yes/no answer


• Storage under sub-optimal conditions - since defined conditions may not be available, the device has to be stable under extreme conditions concerning temperature, pH or humidity for example


• Multiplexing - covering a broad spectrum of possible diseases in only one device allows for a differential diagnosis with the need for only one sample of the patient

The Field of Diagnostics

Medical diagnostics comprise the whole process of tracing the source leading to a patient's symptoms. Usually, it is understood as the identification of a pathogen or a malfunction responsible for the illness.
To achieve an efficient disease treatment, clinical diagnostics are mostly divided into the following steps:

• After interviewing the patient and considering his medical history, risk factors and current problems, a clinician proposes a certain differential diagnosis - thus pre-limiting the spectrum of possible diseases.
  
  
• This is mostly followed by the performance of diagnostic tests (usually in a central laboratory) to confirm the differential diagnosis and to clearly identify - or at least further limit - possible causes of the symptoms.
  
  
• Finally, this leads to a treatment consisting of medication, surgery, hospitalization or discharge.
  

Interest in the so called Point-Of-Care (POC) diagnostics increased dramatically ^{2) 3)} in the last decades. POC diagnostics (or bedside diagnostics) are diagnostic tests that can be performed directly at a patient's site or even bedside. The output of such a test is immediately available, circumventing the usually necessary sending of samples to external labs. The term POC encompasses many possible end-use settings outside of a centralized testing facility like emergency settings, regional health clinics, medical practices as well as home or mobile use.
These tests are devices that are present in everyday life, such as AB0-testing, blood glucose testing, blood gas and electrolytes analysis, pregnancy testing and cholesterol screening.
For the near future an increase in the amount of products for POC diagnostic is predicted ⁴⁾ , thereby confirming the need for such applications. In the face of aging populations, spreading of infectious diseases especially in the developing world, biohazard threats and increasing numbers of autoimmune diseases and allergies in the developed world ^{5) 6)} , POC tests become inevitable.

Current Diagnostic Methods and Limitations

The four most common centralized laboratory techniques are blood chemistry, immunoassays, nucleic-acid amplification tests and flow cytometry ²⁾ . As the DiaCHIP is an immunodiagnostic method, we will focus on this part of diagnostics and compare it to commonly applied methods in today's clinics and labs. Immunodiagnostics are based on antigen-antibody interactions, which might be present within the body fluids of a patient. By detecting and identifying key proteins within a patient's sample like blood or urine, these tests enable to distinguish between major classes of diseases like infectious, metabolic, or cardiovascular diseases or cancer ³⁾ . Immunodiagnostics are realized by immunoassays, which summarize a wide range of formats, allowing quantification and monitoring of small molecules, large proteins and even whole pathogens ²⁾ . Three prominent examples of immunoassays are lateral flow tests, ELISAs and miniaturized immunoassays (microarrays).

ELISA - sensitive but time consuming, no multiplexing:

The commonly used ELISA (enzyme linked immunosorbent assay) only provides a limited capacity for multiplexing as only one specific disease can per detected per well. It takes several hours and large amounts of sample material as well as antibodies (0.05-1.2 µg antibody per well ⁷⁾ ) to test for all the possible diseases fitting the patient's symptoms.

Lateral flow test - simple and rapid, no multiplexing, limited sensitivity

Lateral flow tests are much faster to perform but can only detect one molecule of interest at once. Moreover, they are known to perform rather poorly in terms of sensitivity