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Norwalk virus expression detecting kit and its special primer and probe

A norovirus and detection kit technology, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, etc., can solve the problems of time-consuming electrophoresis, low specificity, and prone to false positives, etc.

Inactive Publication Date: 2006-08-09
中国疾病预防控制中心营养与食品安全所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Overcome many problems of conventional PCR detection technology, such as time-consuming product electrophoresis, ethidium bromide toxicity, prone to false positives, low specificity and other problems

Method used

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  • Norwalk virus expression detecting kit and its special primer and probe
  • Norwalk virus expression detecting kit and its special primer and probe
  • Norwalk virus expression detecting kit and its special primer and probe

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0117] Example 1. Preparation and application of Norwalk virus expression detection kit

[0118] 1. Preparation of kits

[0119] (1), materials and equipment

[0120] PEG6000 (imported packaging, chemically pure) was purchased from Xilong Chemical Factory, Shantou City, Guangdong; NaCl (analytical pure) was purchased from China Pharmaceutical Group Shanghai Chemical Reagent Company; 2 HPO 4 12H2O (analytical pure) was purchased from Shanghai Chemical Reagent Company of China Pharmaceutical Group; 2 PO 4 (analytical pure) was purchased from China National Pharmaceutical Group Shanghai Chemical Reagent Company; KCI (analytical pure) was purchased from China National Pharmaceutical Group Shanghai Chemical Reagent Company; Trizol reagent was purchased from Invitrogen; chloroform (analytical pure) was purchased from Sinopharm Chemical Reagent Co., Ltd.; Isopropanol (analytical pure) was purchased from Shanghai Wulian Chemical Factory; absolute ethanol (analytical pure) was purc...

Embodiment 2

[0183] Embodiment 2. The application of kit

[0184] 1. Specimen detection

[0185] 60 parts of oyster samples, 20 parts of stools from patients with diarrhea, and 10 parts of normal people's stools; apply the kit of the present invention to carry out sample pretreatment and virus RNA extraction, take 8 μ l RNA, carry out reverse transcription in 20 μ l total reaction system, use detection The upstream and downstream primers and probes were amplified by PCR on the 7000 quantitative PCR instrument of ABI Company (GG I and GG II types were detected simultaneously), and the standard curve was made by using the standard products at the same time, and the results were processed by the corresponding software to calculate the virus RNA of the test specimen of expression.

[0186] 2. Experimental results

[0187] 1. GG type I: standard test results see figure 1 , the standard curve see figure 2 .

[0188] 2. GG II type: For the test results of standard products, see image 3 , ...

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Abstract

This invention relates to a test reagent box for the expression volume of Norwalk viruses, its special primers and probes, in which, said box includes former process solution A, B, C and D of a sample, a RNA extraction reagent, water without RNA enzymes, a random primer, an inverse transcription buffer solution, dNTP mixed subject, 0.1 MDTT, inverse transcripted enzyme storage solution, PCR reaction solution of GGI and GGII, probes of GGI and GGII, UNG enzymes of GGI and GGII, standard subjects of GGI and GGII, quality control subjects of GGI and GGII.

Description

technical field [0001] The present invention relates to a norovirus expression detection kit and its special primers and probes, in particular to a detection kit for quantitatively detecting the amount of norovirus in a sample and distinguishing its genotype and The special primers and probes belong to the technical field of virus nucleic acid detection. Background technique [0002] Norovirus (formerly known as Norwalk-like virus), first isolated in 1972 from the stool samples of patients with acute gastroenteritis in Norwalk, Ohio, USA, is an RNA virus with a diameter of 27 μm. Recently, Norovirus (NoV) is considered to be the most important pathogen in the outbreak of foodborne diseases. Most of the people who spread it through human-to-human transmission are the second and third generations of patients, and the initial incidents are mostly caused by contamination of food and water, especially fresh seafood. The outbreak has become an increasingly serious public health p...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 李蓉程民
Owner 中国疾病预防控制中心营养与食品安全所
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