Abstract: The disclosure belongs to the field of molecular biological detection, particularly belongs to the field of miRNA detection, and more particularly relates to one-step detection of miRNA. The disclosure provides a primer and probe design method for miRNA detection, a detection composition including a primer and probe designed by the primer and probe design method, and a detection kit including the detection composition. By using the primer and the probe designed by the method disclosed in the disclosure, miRNA detection may be carried out by adopting one-step method, and non-specific amplification is avoided; the method has a high sensitivity of 1000 copies/mL with good specificity, and the operation of the method is simpler and safer with more credible results.

Abstract: A composition for improving the detection performance of fluorescent quantitative PCR. The composition comprises bovine serum albumin, sorbitol, ammonium sulfate, formamide, tetramethylammonium chloride, and at least one of dithiothreitol and betaine. The present invention further relates to a qPCR reaction liquid containing the composition and a preparation method therefor. The composition can improve the sensitivity, specificity, and interference resistance of real-time fluorescent quantitative PCR.

Abstract: Provided in the present invention is a composition capable of detecting and typing novel coronavirus 2019-nCoV, coronavirus 229E, coronavirus NL63, coronavirus OC43, coronavirus HKU1, coronavirus MERSr-CoV, and coronavirus SARSr-CoV. At the same time, further provided is a kit comprising the composition and a method for detecting and typing coronaviruses. The composition of the present invention in combination with a fluorescent probe method and a melting curve method can perform simultaneous detection and typing of seven coronaviruses in one tube.

Abstract: A composition for detecting SARS-CoV-2 is provided; moreover, a kit including the composition, the use of the kit, and a method for detecting SARS-CoV-2 are also provided.

Abstract: The present invention relates to the field of molecular biology detection, more particularly to a method for fluorescent quantitative PCR. The method includes: 1) mixing an upstream and downstream primer pair, a fluorescent probe, and a PCR amplification reagent; and 2) carrying out the fluorescent quantitative PCR, where the fluorescent probe has two quenching groups, in which a first quenching group is located at a 3? end and a second quenching group is labeled on a T base and is 10-15 nt apart from the first quenching group. Using the method for fluorescent quantitative PCR, background signals in the fluorescent quantitative PCR can be reduced; furthermore, the sensitivity of PCR can be improved, the occurrence of false negatives in detection can be reduced, and the amplification efficiency of the fluorescent quantitative PCR can also be improved.

Abstract: The present invention relates to the field of viral nucleic acid detection. In particular, the present invention provides a pretreatment method for viral nucleic acid detection. The method includes mixing a pretreatment solution containing a sample with a nucleic acid releasing agent and a qPCR amplification reagent, wherein the pretreatment solution includes Tris-HCl, EDTA-2Na, sodium chloride, a ribonuclease (RNase) inhibitor, and an antibiotic; and the pretreatment solution has a pH of 6.5-8.0.

Abstract: A nucleic acid release agent, a PCR amplification method and a PCR amplification kit are provided. The nucleic acid release agent includes Tris-HCl, sodium chloride, potassium chloride, tween 20, Triton X-100, ethyl phenyl polyethylene glycol and a strong base; wherein the molar concentration of Tris-HCl is 0.5 mM to 500 mM, the molar concentration of sodium chloride is 20 mM to 500 mM, the volume percentage of Tween 20 is 0.1% to 2%, the volume percentage of Triton X-100 is 0.1% to 3%, the volume percentage of ethyl phenyl polyethylene glycol is 0.1% to 3%, the mass concentration of potassium chloride is 5 mg/mL to 8 mg/mL and the mass concentration of the strong base is 2 mg/mL to 50 mg/mL.