Fluorescent Quantitiaive PCR system

The Fluorescent Quantitative PCR System utilizes real-time fluorescent detection methods for the analysis of PCR template amplification. Real-time PCR/qPCR tests have emerged as the preferred method for determining and quantifying nucleic acids in a variety of biological samples. It is used in several applications, including the analysis of foodborne pathogens, cancer, and hereditary disease. 

The system includes a thermal cycler equipped with an optical detection module. It measures the fluorescence signal produced during each amplification cycle when the fluorophore binds to the target sequence.

The Fluorescent Quantitative PCR System has several scanning models, such as whole block scanning, formulated line scanning mode, and 96-well double-color scanning, which only takes 5.5 seconds. It enables fluorescence detection across 6 channels with no crosstalk between the different channels. 

Principle 

The principle of qPCR is to detect the accumulation of PCR amplicons in a sample. Fluorescence emission from fluorescent DNA-binding dyes or target-specific fluorescently labeled primers or probes is measured in “real time.”

Features 

• The system has an excitation wavelength between 300 and 800 nm and a detection wavelength between 500 and 800 nm. 
• It has a module temperature range of 4 – 105°C.
• The temperature can be controlled with two temperature control modes (block mode and tube mode).
• It has a gradient temperature range of 1 – 36°C.
• The hot lid temperature range is 30 – 110°C.
• It has a maximum of 20 segments for each program with a maximum of 99 cycles. 
• The fluorescence intense detection repeatability is 5%. 
• The system has various special functions, including absolute quantitative automatic analysis, relative quantification, melting curve analysis, SNP Analysis, HRM multi-channel crosstalk calibration, 6 independent temperature zones, background correction, customized parameters, auto gain, etc. 
• The apparatus weighs 55 kg and measures 410 × 386 × 352 mm. 

Components of the System 

The Fluorescent Quantitative PCR System contains the following components that are present in real-time PCR detection systems:

• Reaction module — For each round of DNA amplification, samples are heated and cooled to exact temperatures to induce nucleotide denaturation, annealing, and then polymerase-mediated extension.
• Optical detection system — Measurement of the fluorescence intensity of each PCR reaction in the presence of a fluorescent reporter, such as a DNA-binding dye or labeled probe, allows the identification of the presence of a target of interest within an experimental sample.
• Instrument software — Real-time PCR detection systems are often managed by an attached computer that runs specialized software that launches and monitors runs before assisting in interpreting the data.

Applications 

• For the rapid prenatal diagnosis of fetal aneuploidies 

The fluorescent Quantitative PCR System can be used for the rapid prenatal diagnosis of fetal aneuploidies via chorionic villus sampling. A study conducted by Shin et al. (2016) examined whether fluorescent quantitative PCR could rapidly diagnose fetal aneuploidies by reviewing the medical records of 383 pregnant females who underwent chorionic villus sampling. The results indicated that 18.9% of the cases were identified as trisomy 21, 18, or 13, and 1.0% of the cases were suspected to be mosaicism. The fluorescent quantitative PCR results for common autosomal trisomies were consistent with LTC results, with no false-positive findings. Therefore, fluorescent quantitative PCR can effectively provide quick prenatal screening for common chromosomal trisomies at an earlier stage of pregnancy. 

• For high-throughput screening of pathogens 

The Fluorescent quantitative PCR system can be used to detect foodborne pathogens. It has several advantages over standard culturing methods, including the lesser time required to obtain results, and it offers greater specificity. A study conducted by Reekmans, Stevens, Vervust, and De (2009) developed a PCR assay to detect Bacillus cereus in gelatin without performing an enrichment step before the assay. The results indicated that the PCR assay reduced the detection time from 2 days to 2 hours compared to the standard method. Moreover, no significant difference was observed between the performance of the PCR assay and the standard method. Furthermore, the analysis costs between both methods were similar. 

Strengths 

The Fluorescent Quantitative PCR System is a time-efficient and cost-efficient method for determining and quantifying nucleic acids in various biological samples. It is used in various fields, including medical, biological, veterinary, biotechnology, and forensic research. The system includes various scanning methods that perform scanning within only 5.5 seconds. The system is also equipped with an automatic hot lid that opens and closes automatically, preventing the evaporation of the reagents. The LED excitation light included with the system has a super long service life. The wells have unique bottom detection, which is compatible with reaction volume down to 5μl.

Summary 

• The Fluorescent Quantitative PCR System automatically detects and measures the number of nucleic acids in the sample using the PCR technique. 
• The Fluorescent Quantitative PCR System has several scanning models. It only takes 5.5 seconds to scan the sample. 
• It enables fluorescence detection across 6 channels with no crosstalk between the different channels. 
• The system has various special functions, including absolute quantitative automatic analysis, relative quantification, melting curve analysis, SNP Analysis, HRM multi-channel crosstalk calibration, 6 independent temperature zones, background correction, customized parameters, auto gain, etc. 
• It is used for various applications, including analyzing pathogens, cancer, and hereditary disease. 

References

Shin, Y. J., Chung, J. H., Kim, D. J., Ryu, H. M., Kim, M. Y., Han, J. Y., & Choi, J. S. (2016). Quantitative fluorescent polymerase chain reaction for rapid prenatal diagnosis of fetal aneuploidies in chorionic villus sampling in a single institution. Obstetrics & gynecology science, 59(6), 444–453. https://doi.org/10.5468/ogs.2016.59.6.444

Reekmans, R., Stevens, P., Vervust, T., & De Vos, P. (2009). An alternative real-time PCR method to detect the Bacillus cereus group in naturally contaminated food gelatine: a comparison study. Letters in applied microbiology, 48(1), 97–104. https://doi.org/10.1111/j.1472-765X.2008.02495.x

Löfström, C., Josefsen, M. H., Hansen, T., Søndergaard, M. S. R., & Hoorfar, J. (2015). Fluorescence-based real-time quantitative polymerase chain reaction (qPCR) technologies for high throughput screening of pathogens. High throughput screening for food safety assessment (pp. 219-248). Woodhead Publishing. doi:10.1586/14737159.5.4.493