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Herpes virus type I PCR (polymerase chain reaction) fluorescence quantitative rapid test kit and method

A detection kit and fluorescence quantitative technology, applied in the direction of microorganism-based methods, microorganism measurement/inspection, biochemical equipment and methods, etc., can solve the problems of poor sensitivity, achieve convenient operation, high sensitivity and specificity, and ensure credibility effect

Active Publication Date: 2012-02-01
泰普生物科学(中国)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Latex agglutination test (LA) is to combine specific antibodies with latex particles to form a complex. When HSVAg exists, an antigen-antibody-latex polymer will be formed, and agglutination can be seen with the naked eye. The problem is that the sensitivity is poor

Method used

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  • Herpes virus type I PCR (polymerase chain reaction) fluorescence quantitative rapid test kit and method
  • Herpes virus type I PCR (polymerase chain reaction) fluorescence quantitative rapid test kit and method
  • Herpes virus type I PCR (polymerase chain reaction) fluorescence quantitative rapid test kit and method

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Experimental program
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Effect test

Embodiment 1

[0034] Table 1: Kit Composition

[0035]

[0036]

[0037] Preparation of basic reagents:

[0038] 1.1 Tris: analytically pure, product from a qualified supplier, content 99.7%, infrared qualified, pH (5% water) 10.3-10.9, moisture 0.3%, melting point 167-171°C, qualified absorption system, highest content of impurities qualified.

[0039] 1.2 NaOH: Analytical pure, the product of Beijing Chemical Reagent Factory or a qualified supplier.

[0040] 1.3TritonX100: brown oily liquid, soluble in cold water, and has strong foaming properties.

[0041] 1.4 EDTA: analytically pure, a product from qualified suppliers, white crystalline powder, soluble in water, acidic in solution, insoluble in alcohol, content not less than 99.5%, qualified in aqueous solution reaction and complexing force test , the highest impurity content is qualified.

[0042]1.5 Purified water: use pure water from Taipu Bioscience Co., Ltd., and then treat it with a Milli-Q Biocel pure water machine from...

Embodiment 2

[0056] Embodiment 2 The use of kit of the present invention

[0057] 1. Reagent preparation (reagent preparation area)

[0058] 1. Take out the DNA extraction solution and set aside.

[0059] 2. After determining the number n of reaction tubes to be performed (number of samples + negative + strong positive quality control + weak positive quality control + 4 quantitative reference substances), take out the HSV-I PCR reaction solution and mix n×44μl HSV- I-PCR reaction solution, add n×1μl DNA polymerase system into a centrifuge tube and vortex to mix well. After instantaneous centrifugation, dispense 45μl into each PCR reaction tube, cover the tube cap and transfer to the sample addition area, avoiding Put it in the refrigerator at 4°C for later use.

[0060] 3. Transfer the reference substance and quantitative reference substance to the sample processing area, and put them in a 4°C refrigerator for later use.

[0061] 2. Sample processing (sample processing area)

[0062] 1...

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Abstract

The invention aims at providing a herpes virus type I PCR (polymerase chain reaction) fluorescence quantitative rapid test kit which can detect the specific nucleic acid sequence of pure herpes virus type I in a clinical sample and further achieve the purpose of rapidly judging the existence of the pure herpes virus type I. In order to realize the purpose, the technical scheme of the invention isas follows: the herpes virus type I PCR fluorescence quantitative rapid test kit provided by the invention comprises a DNA (deoxyribonucleic acid) extraction solution, a PCR reaction solution, a DNA polymerase, a positive quality control product, a weak positive quality control product, a negative quality control product and a quantitative reference product, wherein the PCR reaction solution comprises primers and a fluorescent probe, and the primers are divided into an upstream primer and a downstream primer. The herpes virus type I PCR fluorescence quantitative rapid test kit has the beneficial effect of filling in the blank of a fluorescence PCR kit for clinically detecting the pure herpes virus type I (HSV I) in China. Furthermore, a Taqman core technology platform and an arabidopsis thaliana internal control system are utilized for detecting the pure herpes virus type I (HSV I), so that the herpes virus type I PCR fluorescence quantitative rapid test kit has the advantages of highsensitivity, high specificity, stability, timeliness, convenience in operation and the like.

Description

technical field [0001] The invention relates to a disease pathogen gene detection technology, in particular to a herpes virus type I PCR fluorescence quantitative rapid detection kit and method. Background technique [0002] Herpes simplex virus (herpes simplex virus) belongs to the herpesviridae alpha virus subfamily, and the size of the virus plasmid is about 180 nanometers. Human herpes simplex virus is divided into two types, namely herpes simplex virus type I (HSV-I) and herpes simplex virus type II (HSV-II). Type I mainly causes infections of the skin, mucous membranes (oral mucosa) and organs (brain) other than the genitals. HSV-I and HSV-II are prone to cross-infection. HSV infection is due to person-to-person contact. From four months to several years after the occurrence, the number of infected people can reach 50-90% of the total population. Herpes, Kaposi's disease, etc., are sometimes the cause of meningitis and encephalitis. Oral herpes is generally easier...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12R1/93
CPCC12Q1/705
Inventor 何东华姚雪
Owner 泰普生物科学(中国)有限公司
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