QPCR Q: What is QPCR?A: The full name of QPCR in English is Real-time Quantitative PCR Detecting System. i.e. Real-time Fluorescence Quantitative Nucleic Acid Amplification Detection System, also known as Real-time Quantitative Gene Amplification Fluorescence Detection System, or QPCR for short.Q: What is the relationship between QPCR, DNA Detection and PCR? QPCR, DNA testing, and PCR? A: Polymerase Chain Reaction (Polymerase Chain Reaction referred to as PCR) principle by virtue of the sensitive, specific, rapid features won the Nobel Prize in Chemistry in 93. Because of its unique advantages in the detection of pathogens, the developed countries in the related methods and instruments in the research and development is very fast, to become the mainstream of molecular biology diagnosis, and is still in the forefront of academia and application: the first generation of products: gene amplification thermal cycler (DNA Thermal Cycler) + electrophoresis + UV analyzer + qualitative reagents, constituting the qualitative detection of PCR. The second generation of products: Gene Amplification Thermal Cycler + Fluorometer + End-point quantitative reagents, constituting PCR-DNA end-point quantitative PCR detection, which is also divided into end-point ELISA-PCR and end-point Fluoror-quantitative PCR. PCR). The third generation product: real-time quantitative QPCR instrument + real-time fluorescence quantification reagents, constituting QPCR-DAN/RNA real-time fluorescence quantitative detection. Q: What are the characteristics of QPCR compared to previous methods? A: QPCR has at least the following characteristics: fewer instruments are used, only one instrument is used. The detection time is short, only 45 minutes to 1 hour and 10 minutes (reagents vary), while qualitative PCR takes 3-4 hours, enzyme immunoassay endpoint quantification takes 6-8 hours, and fluorescence endpoint quantification takes 2-3 hours. Extremely simple operation: after pre-treatment, the sample can be inserted into the instrument for one hour and the report can be made on the computer without opening the cap and moving the sample (the previous method) to avoid contamination. Accurate results: Qualitative PCR can only characterize the results, which is very rough, and the endpoint quantitative PCR can only detect the fluorescence at the end of 40 thermal cycles, and the measured fluorescence is saturated, which leads to the lack of accuracy of quantification, and belongs to semi-quantitative state. The real-time fluorescence QPCR continuously detects the change of fluorescence value of each sample at each moment of amplification, as shown in Fig. 1: Fig. 1 Xi'an Tianlong TL988 Actual Curve 1 Dynamic range of detection: 10~1,000,000,000 DNA copies/ml Detection accuracy: 0.1 RLU Discrimination rate: 99.7% for 5,000 and 10,000 template copies. The results are summarized in the following table. Q: What is the technical advancement, price and application status of QPCR? A: Real-time fluorescence QPCR is the most advanced nucleic acid molecular diagnostic technology used in the clinic in today's world, which is recognized and respected by the U.S. FDA. The varieties of QPCR reagents that have been approved by the U.S. FDA and have obtained the license for clinical application are: HBV Hepatitis B Virus DNA, HCV Hepatitis C Virus RNA, HIV AIDS Virus RNA, Chlamydi trachomatis (CT), Sexually Transmitted Diseases (STD), Chlamydia trachomatis (C). Sexually Transmitted Diseases, Chlamydia trachomatis, Neiserria gonorrhea (NG), Sexually Transmitted Diseases, Diplococcus gonorrhoeae, Cytomegalovirus (CMV), Sexually Transmitted Diseases, Cytomegalovirus, Mycobacterium tuberculosis (Mtb), Respiratory Diseases, Mycobacterium tuberculosis, and so on. Real-time fluorescence PCR instrument research and development technology is difficult, the threshold is high, only a limited number of well-known companies in developed countries to produce, and the price is expensive: such as the United States ABI/7700-1.2 million dollars, the United States ABI/5700-500,000 yuan, Switzerland, Roche/Lightcycler-700,000 yuan, the United States Stratagene new product 2005 MX3005P-800,000 yuan It can be seen that, in the view of the patient, which hospital applies the QPCR technology is a symbol of advanced diagnostic technology, also in developed countries. Domestic Instruments: Domestic instruments produced by no more than five units, including: Japan's wholly-owned Hangzhou Bozhi real-time fluorescence QPCR instrument (33 holes) Xi'an Tianlong Technology Co., Ltd. TL988 real-time fluorescence QPCR instrument (48 holes) The above products have obtained the national SFDA certification, the offer are more than 450,000 yuan. Domestic reagents: a number of companies produce real-time fluorescence QPCR reagent kits, the price is comparable with the endpoint method reagents, and complete varieties, cheap, easy to buy: Hepatitis B HBV-DNA, Hepatitis C HCV-RNA, AIDS AIDSHIV-RNA, sexually transmitted diseases: NG gonorrhea, Chlamydia trachomatis CT, Mycoplasma urealyticum UU, herpes virus HSV, human papilloma, HPV, eutrophication, euthyroidism, and other diseases, and the price of real-time fluorescence QPCR kits. HPV, Eugenics Series: Cytomegalovirus CMV, Toxoplasma gondii COX, etc. Editor's word Open classification: virus, DNA, PCR, molecular biology, real-time fluorescence quantitative PCR instrument
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