Gene detection reagent kit for SARS virus and its detection method

A SARS virus and genetic detection technology, applied in the field of genetic detection kits, can solve problems such as unfavorable control of the epidemic

Inactive Publication Date: 2004-01-21
THE FIRST AFFILIATED HOSPITAL ZHEJIANG UNIV COLLEGE OF MEDICINE +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the current clinical diagnostic criteria can only be diagnosed after the SARS patient becomes ill or even becomes seriously ill.
This is obviously not conducive to controlling the epidemic

Method used

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  • Gene detection reagent kit for SARS virus and its detection method
  • Gene detection reagent kit for SARS virus and its detection method
  • Gene detection reagent kit for SARS virus and its detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0081] 优选SARS病毒的基因检测试剂盒,包括SARS病毒特异性核酸定量标准品即SARS的部分序列:5'-TACCGTAGACTCATCTCTATGATGGGTTTCAAAATGAATTACCAAGTCAATGGTTACCCTAATATGTTTATCACCCGCGAAGAAGCTATTCGTCACGTTCGTGCGTGGATTGGCTTTGATGTAGAGGGCTGTCATGCAACTAGAGATGCTGTGGGTACTAACCTACCTCTCCAGCTAGGATTTTCTACAGGTGTTAACTTAGTAGCTGTACCGACTGGTTATGTTGACACTGAAAATAACACAGAATTCACCAGAGTTAATGCAAAACCTCCACCAGGTGACCAGTTTAAACATCTTATACC-3'及(1)荧光扩增检测试剂:由下列组分以蒸馏水为溶剂混合 Made: -step method RT-PCR buffer is: 50mM potassium chloride (KCl), 10mM Tris-Cl, 25mM magnesium chloride (MgCl2), 0.1% polyethylene glycol 6000, 0.1% 1,4-dimercaptothreose Alcohol (DTT), 1% bovine serum albumin (BSA) deoxynucleoside triphosphate (dNTP mixture) is: 200 μM dATP, 200 μM dCTP, 200 μM dGTP, 200 μM dTTP, 50 μM dUTP Specific primer S1 0.6 μM Specific primer S2 0.6 μM Specific probe S3 0.2μM, R is FAM (carboxy fluorescent yellow), Q is TAMARA (tetramethyl-6 carboxyrhodamine), (2) DNA polymerase (Taq): select Ampli TaqR DNA polymerase 2.0 enzyme activity unit / reaction (U) (3) uracil gly...

Embodiment 2

[0093] The gene detection kit of preferred SARS virus, comprises SARS virus-specific nucleic acid quantification standard item as embodiment 1, also comprises: (1) fluorescent amplification detection reagent: be mixed by following components with distilled water as solvent: one-step RT -PCR buffer is: 500mM potassium chloride (KCl), 100mM Tris-Cl, 25mM magnesium chloride (MgCl2), 0.1% polyethylene glycol 6000, 0.1% 1,4-dimercaptothreitol (DTT), 1% Bovine Serum Albumin (BSA). Deoxynucleoside triphosphate (dNTP mixture) is: dATP:dCTP:dGTP:dTTP:dUTP=4:4:4:4:1; the concentration used is 100 μM specific primer S1 0.45 μM specific primer S2 0.55 μM specific probe Needle S3 0.2μM, R is FAM (carboxy fluorescent yellow), Q is TAMARA (tetramethyl-6 carboxyrhodamine) (2) DNA polymerase (Taq): Ampli TaqR DNase is selected, 1.5 enzyme activity units / reaction (U )(3) Uracil glycosylase (UNG): 200 enzyme activity units / reaction (U) (4) reverse transcriptase M-MuLV 200 enzyme activity units / ...

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Abstract

The detection process of SARS virus of the gene detection reagent kit with rationing standard specific SARS virus nucleic acid as reference includes RNA extraction, PCR amplification and fluorescent detection. The venous blood sample, gargled liquid or respiratory tract secretion of the patient is used as the analysis sample directly, RNA of the sample is extracted as the nucleic acid template, and the template is used in fluorescent PCR amplification. The used fluorescent amplification detecting reagents includes one-step process RT-PCR buffering liquid, deoxynucleoside triphosphate (dNTP) mixture, specific amplification primer and specific probe. The method of the present invention is fast and convenient in detecting SARS virus.

Description

1. Technical field [0001] The invention relates to a gene detection kit and a gene detection method for detecting whether a sample contains SARS virus by using the kit. 2. Background technology [0002] Atypical pneumonia is an acute respiratory infectious disease, which is called severe acute respiratory syndrome (Severe Acute Respiratory Syndrome, SARS) abroad, or SARS for short. It is mainly transmitted through close-range air droplets and close contact. It has an acute onset and rapid spread, and the case fatality rate exceeds 5%. Since the first case appeared in Guangdong Province of my country in November 2002, nearly 30 countries and regions in the world have been involved in just half a year, and the number of patients worldwide has reached more than 6,000, and the number of cases is still increasing. Because it is transmitted through close-range air droplets and close contact, there is currently no vaccine or specific drug, and most people are not immune to SARS. C...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/25C12Q1/68
Inventor 李兰娟姜汉卿沃健儿吴南屏邵俊斌麻静明
Owner THE FIRST AFFILIATED HOSPITAL ZHEJIANG UNIV COLLEGE OF MEDICINE
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