The field of real-time PCR (RT-PCR) is experiencing a rapid and remarkable expansion. The global demand for faster and high-throughput testing was further fueled by the SARS-CoV-2 pandemic. As a result, improved RT-PCR methods have emerged, characterized by reduced reagent consumption, enhanced reproducibility, and heightened precision. These advanced machines automate the workflow and incorporate real-time monitoring, offering unparalleled accuracy and customizable options.
PCR: A Breakthrough in Infectious Disease Testing
Polymerase chain reaction (PCR) represents a pivotal breakthrough in biotechnology and has become an indispensable tool in the healthcare industry’s arsenal of test kits and solutions. In this article, we will delve deeper into the role of RT-PCR in infectious disease testing.
PCR vs. qPCR in Infectious Disease Testing
PCR, including qPCR/RT-PCR, serves to amplify DNA for various applications. PCR is highly effective for amplifying DNA from bacteria, viruses, and other pathogenic organisms. However, when it comes to amplifying RNA and converting it into DNA for amplification, RT-PCR/qPCR processes are more suitable.
Distinguishing PCR Testing and RT-PCR in Infectious Disease Diagnosis
PCR testing employs a qualitative approach, determining whether a specific DNA sequence is present or absent. On the other hand, RT-PCR utilizes a quantitative method, providing information about the quantity of viral DNA present. PCR is commonly utilized for diagnosing diseases such as African swine fever, Ebola, and foot-and-mouth disease.
Unraveling the RT-PCR Testing Process
During RT-PCR testing, the machine utilizes precise heating and cooling cycles to trigger chemical reactions, resulting in the creation of identical copies of targeted segments of viral DNA. These copies double in number with each cycle, enabling efficient amplification.
The RT-PCR Process: An Insight into the Steps
In the initial step of phase one, the samples are subjected to a temperature of 50.0°C for two minutes to establish temperature equilibrium and prepare the samples for the subsequent reactions.
The second step involves raising the temperature to 95.0°C for ten minutes to activate the polymerase in the PCR mix.
Following this, phase two initiates the amplification process:
- Denaturing phase: The temperature is set to 95.0°C for 15 seconds.
- Annealing and elongation phase: The temperature is maintained at 60.0°C for 1 minute.
The machine repeats phase two for 40 cycles, resulting in the generation of multiple copies of the DNA strands. Fluorescent markers are attached to each new DNA copy, and a fluorescent dye is released by the marker during amplification. The machine’s computer tracks the fluorescence levels in real-time, displaying the data on the monitor. Once the fluorescence surpasses a predetermined threshold, the presence of the virus is confirmed.
Realizing the Potential of RT-PCR in Infectious Disease Testing
Real-time PCR has revolutionized the field of infectious disease testing, providing rapid, accurate, and quantitative results. By harnessing the power of this advanced technology, healthcare professionals can swiftly and precisely diagnose various infectious diseases, aiding in effective management and control.
Real-time PCR (RT-PCR) has found extensive clinical application in the diagnosis of viral diseases, with its importance highlighted during the COVID-19 pandemic. Rapid detection facilitated efficient management of limited isolation beds and critical care for patients during the early stages of the outbreak.
RT-PCR Testing for COVID-19
RT-PCR testing is employed to detect the presence of SARS-CoV-2 viral nucleic acids or proteins, providing high specificity for the viral nucleic acid. However, it is important to note that this test cannot differentiate between a viable and non-viable virus. Therefore, it cannot determine whether an individual is currently infectious, impacting isolation requirements.
RT-PCR Thermal Cyclers: Powering Precision and Performance
Leading brands of RT-PCR Thermal Cyclers, such as QIAquant instruments and qTOWER³, offer exceptional temperature control precision with their thermal blocks, regardless of the number of samples being processed. These instruments ensure fast and sensitive detection through high-performance optics.
Homogeneous Illumination and Excitation
QIAquant instruments and qTOWER³ provide homogenous illumination and excitation for each sample, maximizing the accuracy of results. These machines employ proven filter modules that offer remarkable flexibility with each cycle. The modules can be expanded and configured to enable up to six-fold multiplexing, enhancing testing capabilities.
QIAquant instruments, equipped with rapid heating and cooling ramp rates and high-quality fiber-optic shuttles, significantly reduce processing times by completing 40 cycles in under 30 minutes. The qTOWER³ 84, featuring a 16-fiber high-performance optic, delivers results for a full 348-well plate in just six seconds, irrespective of the number of filters used.
By harnessing the power of advanced thermal cyclers like QIAquant and qTOWER³, laboratories can ensure precise and efficient real-time PCR testing, enabling faster and more accurate diagnoses for various viral diseases.
Discover Your Trusted Partner for Laboratory Supplies
In this article, we explored the significance of real-time PCR in the field of biomedicine. When it comes to reliable laboratory supplies, look no further than Complete Laboratory Systems. CLS offers a comprehensive range of products and equipment to meet your laboratory needs.
At Complete Laboratory Systems, you can find a wide selection of CLIA Waived, moderate, and high complexity lab tests, ensuring that you have access to the necessary tools and equipment for accurate and efficient testing. Whether you require specific reagents, instruments, or other lab essentials, we have you covered.
Experience the convenience of shopping for your lab products with Complete Laboratory Systems. View the essential RT-PCR and molecular diagnostics category.