78 results about "Uracil-DNA glycosylase" patented technology

The invention discloses a fluorescence quantitative PCR detection kit of hepatitis B virus and an application thereof. The kit is composed of the following independent components: DNA extraction solution I, DNA extraction solution II, DNA extraction solution III, DNA extraction solution IV, positive control interior label, PCR reaction liquid, probe HBV-SP, enzyme mixed liquor containing heat resistant DNA polyase and uracil DNA glycosylase, quantitative hepatitis B virus reference material, hepatitis B virus positive control serum and hepatitis B virus negative control serum, wherein DNA extraction solution I contains 0.2-1.0% of lauryl sodium sulphate (mass / volume), 1.0-4.0% of Triton (volume / volume) and 0.2-1.0mol / L of guanidinium isothiocyanate; DNA extraction solution II contains 100-300mmol / L of 4-HEPES, 100-300mmol / L of sodium chloride with pH of 6.5+ / -0.2 and 100-400 mu g / ml of magnetic beads; DNA extraction solution III contains 0.1-1.0% of Triton (volume / volume) and 100-300mmol / L of sodium chloride; DNA extraction solution IV contains mineral oil. The fluorescence quantitative PCR detection kit of hepatitis B virus of the invention can be used for detecting the HBV-DNA concentration in samples of serum, blood plasma or latex and the like.

The invention provides a fluorescent polymerase chain reaction (PCR) kit for detecting CYP2C19 genotypes, and belongs to the field of in-vitro nucleic acid testing. The fluorescent PCR kit comprises PCR reaction liquids for detecting CYP2C19*2 and CYP2C19*3 genotypes, Taq DNA polymerase, and uracil-DNA glycosylase, wherein the PCR reaction liquids for detecting the CYP2C19*2 and CYP2C19*3 genotypes respectively comprise PCR amplification primers, minor groove binder (MGB) probes and the like; and nucleotide sequences for detecting the CYP2C19*2 and CYP2C19*3 genotypes are shown as SEQ ID NO:3-4 and SEQ ID NO:5-6 respectively. The kit has high sensitivity and specificity, can monitor the reaction progress in real time, ensures short reaction time, avoids subsequent treatment, can avoid reaction product pollution to the greatest extent, and can replace the traditional protein detection or the common PCR detection to diagnose the CYP2C19 genotypes.

The invention provides a method for parallel determination of the activity of uracil-DNA glycosylase and endonuclease IV, application thereof and a reagent kit. When a target object is UDG, if UDG exists, U basic groups in a hairpin probe are removed, an AP locus is generated, the generated AP locus is cut through tool enzyme Endo IV, and a primer sequence containing free 3' terminal is released and used for initiating subsequent rolling circle amplification reaction (RCA); if UDG does not exist, the 3' terminals are closed in the hairpin probe, the RCA process can not be carried out. When a target object is Endo IV, if Endo IV exists, the RCA process is the same as that in UDG activity detection; if Endo IV does not exist, the 3' terminals are closed in the hairpin probe, and the RCA process can not be conducted. The hairpin probe can be used as a recognition probe of UDG and can also be used for precursor recognition probe of Endo IV, and accordingly design of the probe is simplified. The detection limits of UDG and Endo IV are reduced to 0.00017 U / mL and 0.11 U / mL respectively, and results are better than or the same as those of other label-free fluorescence methods.

The invention relates to a loop-mediated isothermal amplification kit and method for detecting fusarium. The kit comprises loop-mediated isothermal amplification reaction liquid, UNG (uracil-DNA-glycosylase), Bst DNA polymerase, developer and fusarium positive DNA, wherein the reaction liquid contains forward / reverse inner and outer primers, of which the sequences are respectively SEQ ID NO: 1-4. The detection method comprises the steps of fungus DNA extraction, loop-mediated isothermal amplification and development detection. The loop-mediated isothermal amplification primers are designed according to the fusarium gene conserved region sequences, and the fusarium in the sample is detected by loop-mediated isothermal amplification technology. The invention has the advantages of high speed, high sensitivity, high specificity, low cost and the like; and the whole process is not related to toxic reagents, thereby ensuring the safety of operating personnel and environment. Meanwhile, the UNG adopted in the invention can thoroughly eliminate false positive caused by nucleic acid pollution in multiple detection processes.

The invention relates to a PCR detection kit for specific detection of fowl adenovirus group I and a detection method thereof. The detection kit comprises a fowl adenovirus group I PCR reaction liquid, 10*PCR buffer for UNG plus, hot start type DNA polymerase, dU plus dNTP mix, uracil DNA glycosylase, positive quality control serum and negative quality control serum, wherein the fowl adenovirus group I PCR reaction liquid comprises an upstream primer with a concentration of 5 micrometer and the sequence is shown as the specifications as well as a downstream primer with a concentration of 5 micrometer and the sequence is shown as the specifications. When detection is conducted through the PCR detection kit, conditions for a PCR amplified reaction are that UNG treatment is conducted at a temperature of 25 DEG C for 10 min, and predegeneration is conducted at a temperature of 95 DEG C for 2 min; degeneration is conducted at a temperature of 98 DEG C for 10 s, annealing is conducted at a temperature of 55 DEG C, extension is conducted at a temperature of 72 DEG C for 1 min, and all that are conducted for 30 work cycling; extension is conducted at a temperature of 72 DEG C for 10 min. The PCR detection kit for the specific detection of the fowl adenovirus group I has the advantages of being high in sensitivity (still positive when the reaction liquid is diluted to 1:105) and good in repeatability. The PCR detection method for the specific detection of the fowl adenovirus group I is easy, convenient and fast to operate, suitable for early diagnosis of fowl adenovirus group I virus, and can meet the demand for disease prevention and control in time.

The invention provides a fluorescent PCR kit for qualitative detection of HLA-B*1502 gene subtypes, and belongs to the field of in-vitro nucleic acid detection. The kit comprises uracil DNA glycosylase, Taq polymerase, PCR genotyping primers and a fluorescence probe; the PCR genotyping primer sequence is as shown in SEQ ID NO: 3-4 and / or SEQ ID NO: 6-7. The kit provided in the invention has high sensitivity and good specificity, and can monitor reaction process in real time and the reaction time is short; in addition, closed tube operation is performed, and subsequent treatment is not needed, which can maximally avoid the pollution of a reaction product, thus being capable of replacing traditional cell detection.

A promising approach for the therapeutic treatment of brain tumors utilizes replication-competent, neuroattenuated herpes simplex virus-1 (HSV-1) mutants. This approach requires mutation of HSV-1 to eliminate killing of normal, non-dividing cells of the brain (e.g., neurons). The present invention discloses methods for killing malignant brain tumor cells in vivo entails providing replication competent herpes simplex virus vectors to tumor cells. A replication competent herpes simplex virus vector, with defective expression of the gamma 34.5 gene and the uracil DNA glycosylase (UNG) gene, specifically destroys tumor cells, is hypersensitive to anti-viral agents, and is not neurovirulent.