BackPCR in Real-Time – The Latest Technologies in Diagnosing Infections
DNA diagnostics is one of the most modern high-tech analysis methods. DNA tests are widely used in the diagnosis of infectious diseases, making it possible to detect even single microorganisms in the human body.
DNA diagnostics combines several analysis methods, the most common of which is the PCR (polymerase chain reaction) method.
What is PCR?
PCR stands for Polymerase Chain Reaction, PCR diagnostics.
PCR diagnostics is a method of laboratory diagnosis of infectious diseases, in particular, this method is widely used for the diagnosis of STDs.
The PCR test is based on the detection of a small fragment of DNA of the causative agent of the infection that the doctor suspects in the study material. A Small Fragment of DNA is several hundred pairs of DNA bases – building blocks arranged in a strictly defined sequence, and therefore forming a unique pattern. A small fragment is enough for PCR diagnostics of infections since any DNA includes at least several thousand bases.
During the PCR tests, a fragment of the infection DNA is searched for that is specific only to this microorganism. This means that this piece of DNA is Special – it is found only in this microbe (or group of related microbes), but not in any other microbe.
The polymerase chain reaction itself is used to multiply and clone the fragment found: to unambiguously "see" these DNA fragments, there must be at least 1012 of them by the end of the reaction.
The history of the discovery of DNA and the development of the PCR method
Initially, the principle of the polymerase chain reaction (PCR) method was developed by Kary Mullis in 1983. The discovery of PCR was one of the most remarkable developments in the field of molecular biology in the last 20 years, and Kary Mullis was awarded the Nobel Prize in Chemistry in 1993 for the development of the PCR test.
The emergence of the polymerase chain reaction method was preceded by certain achievements in molecular genetics: by that time, the nucleotide sequences of the genomes of a number of microorganisms had already been deciphered and specific ones had been isolated.
Also, the discovery of the unique enzyme DNA polymerase (or taq polymerase) greatly contributed to the emergence of PCR. It is this enzyme that catalyzes and "controls" all processes during the PCR test. The peculiarity of this enzyme is that it is heat-stable and extremely heat-resistant: it withstands heating to boiling point without loss of activity, and its "favorite" temperature regime during operation is 72°C. Many PCR reactions occur almost exclusively at elevated temperatures.
Since the advent of the method, PCR analysis has been gaining more and more popularity. At present, a new PCR technology has been introduced into practical healthcare – Real-Time PCR. Its principal feature is the monitoring and quantitative test of the accumulation of polymerase chain reaction products and the automatic registration and interpretation of the results obtained. This method does not require an electrophoresis step, which avoids errors and false positives associated with contamination and significantly speeds up the result. Real-time PCR uses fluorescently labeled oligonucleotide probes to detect DNA during the amplification process. Real-time PCR allows for a complete test of a sample within 20-60 minutes and is theoretically capable of detecting even a single DNA or RNA molecule in a sample.
OLYMP CDL performs PCR tests on the only 6-channel analyzer Rotor-Gene™ 6000 (Germany) in Kazakhstan. A unique feature of this device is that it allows simultaneous detection of 5 different pathogens! Other laboratories in Astana and nearby regions do not yet have such capabilities.
PCR test material
The material for PCR diagnostics can be:
Epithelial cell scraping (urethral scraping in men and women, scraping from the cervical canal)
Blood, Plasma, Serum
Blood
Biological
fluids (prostate fluid, pleural, cerebrospinal fluid, amniotic fluid, joint fluid, saliva)
urine (the first portion of morning urine is used)
phlegm
mucus
and other biological excretions
How to properly prepare for a PCR test (DNA diagnostics)
The accuracy of the results of laboratory diagnostics depends not only on the experience and professionalism of the laboratory assistant, and the capabilities of this laboratory, but also on whether the patient followed the doctor's recommendations, and how correct his preparation for the test was. There is nothing difficult about proper preparation. When taking a PCR test, doctors recommend following the following rules:
For PCR tests for sexually transmitted infections, it is advisable to refrain from taking antibiotics and medical procedures one month before taking the sample (unless otherwise prescribed by your doctor).
In men, the analysis is carried out in the morning or during the day before urination, it is recommended not to urinate for 4-5 hours before taking the sample.
Women do a PCR test before menstruation or 1-4 days after it ends. Immediately before giving the scraping, it is necessary to abstain from sexual activity, and before taking material from the urethra - from urination for 1-2 hours. On the eve of the examination, women should not conduct douching.
Where can I get PCR tested?
PCR tests can be taken at medical centers that are partners of OLYMP CDL:
Shakhar Medical Center, Abylai Khan Ave. 6/2, phone No.: 35-94-71;
DaNel Medical Center, Auezov Str. 22, phone No.: 39-16-75, 39-16-22;
If you have any questions, you can also get the necessary information from a qualified consultant by phone: 8 (7172) 444-111
