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Real-time PCR

Real-time polymerase chain reaction, sometimes known as qPCR, is a polymerase chain reaction (PCR)-based molecular biology laboratory technique. Instead of detecting amplification at the end of the PCR like in traditional PCR, it monitors the amplification of a targeted DNA molecule while it is happening (i.e., in real time). Real-time PCR can be employed semi-quantitatively (i.e., above/below a specific amount of DNA molecules) (semi-quantitative real-time PCR) and quantitatively (quantitative real-time PCR).
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Real-time PCR and Infectious Diseases

Infectious disorders can transmit from one person to another either directly or indirectly because they are brought on by pathogenic microorganisms such as bacteria, viruses, parasites, or fungus. Unfortunately, even in clinical laboratories, the diagnosis of infectious disorders is directly related to the length of time it takes to identify the pathogen using traditional culture methods, which can take anywhere from hours to days. Benefits of real-time PCR have evolved over the past ten years to become the industry standard for diagnosing infectious illnesses. Infectious disease specialists have championed real-time PCR because of its exceptional sensitivity, specificity, reproducibility, wide dynamic range, and speed of amplification for identifying organisms that cannot be grown in vitro or in situations where existing culture techniques are insensitive and/or require prolonged culture times.


Coronavirus disease 2019, officially named COVID-19 by the World Health Organization (WHO) is a severe acute respiratory syndrome (SARS) caused by the novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Coronaviruses are enveloped, positive-sense, single-stranded RNA viruses, with at least six open reading frames. The coronavirus genome encodes the structural glycoproteins spike (S), membrane (M), envelope (E), and nucleocapsid (N). COVID-19 can be diagnosed in the laboratory using RT-qPCR by detecting SARS-CoV-2 genes in clinical samples collected from suspected patients. For sensitive detection of SARS-CoV-2, a standardised and validated assay with extremely accurate laboratory performance is crucial.


Human monkeypox (MPX) is an emerging zoonotic disease caused by monkeypox virus (MPXV), a double-stranded DNA virus that belongs to the genus Orthopoxvirus within the family Poxviridae, subfamily Chordopoxvirinae. Despite being discovered in captive monkeys in 1958 and humans from the Democratic Republic of the Congo (DRC) in 1970, an MPXV diagnostic test is not frequently used in diagnostic labs. An accurate and rapid real-time PCR method that can be used to diagnose MPXV in humans has thus been developed.


Malaria is still a major issue worldwide. According to the Centers for Disease Control and Prevention, malaria should be considered in the differential diagnosis of febrile patients who have travelled to an area where it is endemic, as well as in all patients who experience fevers of unknown origin, regardless of their travel history. For many laboratories, providing these diagnostic services and maintaining expertise may be difficult. Real-time PCR diagnosis of malaria has advantages over traditional microscopic methods, especially when parasitaemia is low and when dealing with mixed infections. Rapid and accurate species differentiation of Plasmodium is also needed to institute proper therapy, particularly P. falciparum, since it is the species that causes approximately 95% of malaria-related deaths.



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