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Reagent kit for detecting rotavirus nucleic acid

A virus nucleic acid and kit technology, which is applied in the field of kits for detecting rotavirus nucleic acid using real-time fluorescent PCR technology, can solve the problems of lack of sensitivity and difficulty in popularization, and achieve fast detection speed, strong controllability, and high sensitivity Effect

Active Publication Date: 2010-03-03
DAAN GENE CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] 1. Electron microscope observation: Sometimes immunoelectron microscopy and ultrathin section electron microscopy are also used. The role of this method in diagnosis is highly appraised, but due to equipment limitations, it is difficult to popularize
[0006] 2. Enzyme immunosorbent assay: the double sandwich method is the most commonly used detection method at present. WHO lists this method as the standard method for diagnosing RV. Related products are already on the market at home and abroad, but this rapid detection The sensitivity of the method has a relatively large deficiency

Method used

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  • Reagent kit for detecting rotavirus nucleic acid
  • Reagent kit for detecting rotavirus nucleic acid
  • Reagent kit for detecting rotavirus nucleic acid

Examples

Experimental program
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Effect test

Embodiment 1

[0035] Example 1: One-step kit detection method for rotavirus

[0036] (1) Specimen collection, transportation and storage

[0037] Take 500-1000μl of the sample to be tested and the watery diarrhea, put it into a sterile 1.5ml centrifuge tube, and send it directly for testing or store it at -20°C, and the storage period should not exceed six months. Ship sealed in dry ice to avoid freeze-thaw.

[0038] (2) Nucleic acid extraction

[0039] a) Take 100 μl of water sample excreta (if it has been dried, add appropriate amount of pure water or saline to resuspend), place it in a 1.5ml sterilized centrifuge tube, add 200 μl of lysate and 100 μl of chloroform, shake vigorously with an oscillator for 20 seconds Let stand for 3 minutes, then centrifuge at 12,000 rpm for 2 minutes. Yin and Yang quality control products are the same as above.

[0040] b) Carefully take out the colorless upper layer liquid (be careful not to touch the middle flocculent layer), transfer it to a newly ...

Embodiment 2

[0067] Example 2: Use of Quality Controls in Rotavirus Nucleic Acid Detection Kit

[0068] The quality control substances in the rotavirus nucleic acid detection kit include positive quality control substances and negative quality control substances, which are used for quality control in clinical trials, and the operation method is the same as that of samples to be tested.

[0069] The quality control standards are as follows:

[0070] There is no obvious S-type growth in the amplification curve of the negative quality control product;

[0071] The amplification curve of the positive quality control product is S-shaped and the Ct value is less than 30;

[0072] The above requirements must be met in the same experiment at the same time, otherwise, this experiment will be deemed invalid and must be repeated.

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Abstract

The invention relates to a reagent kit for detecting rotavirus nucleic acid, in particular to a reagent kit for detecting rotavirus nucleic acid by using a fluorescence PCR technique. The reagent kitcan be used for qualitatively detecting the rotavirus nucleic acid. Due to the characteristics of high sensitivity, high specificity, simple operation and high detecting speed, the reagent kit can bewidely used for the auxiliary diagnosis of rotavirus infections.

Description

technical field [0001] The invention relates to a rotavirus detection kit, in particular to a kit for detecting rotavirus (Rotavirus) nucleic acid by using real-time fluorescent PCR technology. The kit can be widely used in auxiliary diagnosis of rotavirus infection by performing qualitative detection of rotavirus nucleic acid in samples. Background technique [0002] Rotavirus is the most common pathogen in viral gastroenteritis. Group A rotavirus mainly infects infants and young children. It has been reported that the annual number of cases in developing countries exceeds 130 million, while the death toll exceeds 870,000. Group B rotavirus (also known as adult diarrhea rotavirus) can cause outbreaks of gastroenteritis in young adults. In 1973, Australian scholar Bishop et al first discovered it in ultra-thin sections of the duodenal mucosa of children with acute non-bacterial gastroenteritis, and named it rotavirus in 1974. [WhiteGB, Ashton CI, Roberts C, Parry HE.1974....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68
Inventor 周高英朱坤李明程钢何蕴韶
Owner DAAN GENE CO LTD
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