Where can quantitative real-time PCR be applied?
Overview of qPCR
Quantitative Real-time PCR is a method for measuring the total amount of product after each polymerase chain reaction (PCR) cycle with fluorescent chemicals in a DNA amplification reaction. A method for quantitative analysis of specific DNA sequences in a sample to be tested by an internal or external reference method.
The so-called real-time fluorescence quantitative PCR technology refers to the method of adding fluorophores to the PCR reaction system, using fluorescence signal accumulation to monitor the whole PCR process in real time, and finally quantitative analysis of the unknown template by standard curve.
Where can qPCR be applied?
Due to its powerful advantages, qPCR has a wide range of applications. The method has been around long enough for the research community to demonstrate its reliability and robustness.
Clinical disease diagnosis
Diagnosis and efficacy evaluation of various infectious diseases such as hepatitis, AIDS, avian influenza, tuberculosis, and sexually transmitted diseases; eugenics and prenatal testing for thalassemia, hemophilia, gender dysplasia, mental retardation syndrome, and fetal malformation; tumor marker and tumor gene detection Realize the diagnosis of tumor diseases; genetic testing realizes the diagnosis of genetic diseases.
Animal disease detection
Avian influenza, Newcastle disease, foot-and-mouth disease, swine fever, Salmonella, Escherichia coli, Actinobacillus pleuropneumoniae, parasitic diseases, etc., Bacillus anthracis.
Food safety
Detection of food-derived microorganisms, food allergens, genetically modified organisms, and Enterobacter sakazakii in dairy companies.
Scientific research
Quantitative research on molecular biology related to medicine, agriculture and animal husbandry, and biology.
Application industry
Various medical institutions at all levels, universities and research institutes, CDC, Inspection and Quarantine Bureau, veterinary stations, food companies and dairy factories, etc.
Since qPCR is a real-time quantitative detection of pathogenic pathogen gene nucleic acid, it has unique advantages over immunological methods such as chemiluminescence, time resolution, and protein chips.
The most obvious is the use of qPCR in molecular diagnostics, which is slowly replacing traditional methods. It is used to detect, identify and quantify disease-causing microorganisms (bacteria, viruses and fungi). qPCR reduces manual labor while reducing concerns about contamination and erroneous results. It also allows the processing of large numbers of samples in a shorter time and thus has proven to be an irreplaceable method in diagnostic laboratories. It must be noted, however, that this method only detects the presence of microbial DNA or RNA and does not report on their viability. Therefore, traditional microbiological techniques are still sometimes required.
qPCR is also used to detect and quantify GMOs or perform genotyping. The latter means that different alleles or single nucleotide polymorphisms (SNPs) of the same gene can be detected and can be used as genetic diagnostic or prognostic markers for certain diseases.
A very important application area is gene expression research, which helps us understand biological processes in various fields of biology, microbiology, medicine and other life sciences. A very useful and almost blockbuster combination is genome-wide gene expression screening using DNA microarrays followed by validation of the results using qPCR.