Fluorescent quantitative detection kit for simultaneously detecting human influenza virus and novel coronavirus
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A fluorescent quantitative detection, coronavirus technology, applied in the determination/inspection of microorganisms, resistance to vector-borne diseases, biochemical equipment and methods, etc., can solve problems such as repeated detection, achieve multiple rapid detection, strong specificity , the effect of simple operation
Inactive Publication Date: 2020-10-09
上海同进基因科技有限公司
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It solves the problem of multiple repeated detection...
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Embodiment 1
[0038] Sample to be tested: Throat swab.
[0039] Check the results against Table 3. The specific amplification curve is as follows: figure 1 shown.
[0040] Such as figure 1 , the negative quality control control has no amplification curve, the positive quality control has S-type amplification curves in the four fluorescence channels of FAM, ROX, Cy5, and VIC, and the sample to be tested only has S-type amplification curves in the FAM fluorescence channel. The Ct value is ≤36, so it is determined that the test sample is positive for influenza A.
Embodiment 2
[0042] Sample to be tested: Throat swab.
[0043] Check the results against Table 3. The specific amplification curve is as follows: figure 2 shown.
[0044] Such as figure 2 , the negative quality control control has no amplification curve, the positive quality control has S-type amplification curves in the four fluorescence channels of FAM, ROX, Cy5, and VIC, and the sample to be tested only has S-type amplification curves in the ROX fluorescence channel. The Ct value is less than or equal to 36, so it is determined that the test sample is positive for influenza B.
Embodiment 3
[0046] Sample to be tested: cDNA from a sample of suspected COVID-19 patients.
[0047] Check the results against Table 3. The specific amplification curve is as follows: image 3 shown.
[0048] Such as image 3, the negative quality control has no amplification curve, the positive quality control has S-type amplification curves in the four fluorescence channels of FAM, ROX, Cy5, and VIC, and the samples to be tested have S-type amplification curves in the two fluorescence channels of Cy5 and VIC. The Ct value is less than or equal to 36, so it is determined that the test sample is positive for new coronary pneumonia.
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Abstract
The invention relates to a fluorescent quantitative detection kit for simultaneously detecting human influenza A virus, human influenza B virus and novel coronavirus, and belongs to the technical field of nucleic acid detection. The detection kit specifically comprises four groups of specific primer pairs and probes, a negative quality control material, a positive quality control material and a fluorescent quantitative PCR reaction system, wherein the four groups of specific primer pairs and probes are respectively a pair of specific primers and a probe for detecting influenza A virus and influenza B virus, and two pairs of specific primers and two probes for detecting novel coronavirus; wherein the positive quality control material is an artificially synthesized target sequence, and the negative quality control material is deionized water; wherein the fluorescent quantitative PCR reaction system is composed of components for PCR reaction and reaction conditions. According to the kit disclosed by the invention, the specific primer pair and the probe are designed at a conservative site of a virus sequence; the kit can realize simultaneous detection of influenza A virus, influenza Bvirus and novel coronavirus in a single tube, has strong detection specificity and high sensitivity, and can accurately and rapidly distinguish influenza from new coronapneumonia patients from people,so as to realize early discovery, early isolation and early treatment. The kit has important application value in the fields of disease monitoring, clinical diagnosis and the like.
Description
technical field [0001] The invention belongs to the technical field of nucleic acid detection, in particular to a fluorescent quantitative detection kit for simultaneously detecting human influenza virus and novel coronavirus. Background technique [0002] Novel coronavirus (2019 Novel coronavirus, 2019-nCoV) is a positive-sense single-stranded RNA coronavirus, a new strain of coronavirus that has never been found in humans before. Common signs after a person is infected with the new coronavirus include respiratory symptoms, fever, cough, shortness of breath, and dyspnea. In more severe cases, infection can lead to pneumonia, severe acute respiratory syndrome, kidney failure, and even death. Epidemiological investigations show that most cases can be traced to close contacts with confirmed cases. Confirmed cases need to have positive results of etiological evidence, that is, real-time fluorescent PCR detection of new coronavirus nucleic acid positive. [0003] More and mor...
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