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Methods and compositions for detecting SARS virus and other infectious agents

A technology of viruses and coronaviruses, applied in biochemical equipment and methods, measurement/testing of microorganisms, resistance to vector-borne diseases, etc., can solve the problem that RT-PCR cannot meet the needs of early clinical screening and diagnosis, RT-PCR detection Low rate, expensive and other issues

Inactive Publication Date: 2006-07-12
CAPITALBIO CORP +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, RT-PCR cannot exclude infected patients before SARS virus expression, and the detection rate of RT-PCR is very low
Detection process requires expensive real-time PCR equipment
Therefore, RT-PCR cannot meet the needs of early clinical screening and diagnosis

Method used

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  • Methods and compositions for detecting SARS virus and other infectious agents
  • Methods and compositions for detecting SARS virus and other infectious agents
  • Methods and compositions for detecting SARS virus and other infectious agents

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0227] Example 1. Probe design

[0228] Various genomic sequences of SARS-CoV are available (see Table 22 for example).

[0229] serial number

SARS crown

Virus

Source

Country of submission

Home (region)

GenBank

Acc

N in the sequence

Number of

Genome leader

Degree

Ratio of N

example

SARS_BJ01

Beijing China

China

AY278488

900

28920

3.11％

SARS_BJ02

Beijing China

China

AY278487

300

29430

1.02％

SARS_BJ03

Beijing China

China

AY278490

607

29291

2.07％

SARS_GZ01

Beijing China

China

AY278489

1007

29429

3.42％

SARS_BJ04

Beijing China

China

AY279354

2502

24774

10.10％

SARS_

CUHK-W1

China Hong Kong

China Hong Kong

AY278554

0

29736

0.00％

SARS_HKU-398

49

China Hong Kong

...

Embodiment 2

[0243] Example 2: Pre-processing of blood samples

[0244]The pre-processing of blood samples involves a relatively complicated process. However, considering the relatively low SARS virus concentration reported in serum, the pretreatment described here can effectively enrich lymphocytes from approximately 2 ml of whole blood in order to increase the chance of detection.

[0245] 1. Sample collection and transmission

[0246] 1) Collect samples from patients in the hospital and put them into the first transmission window. Then close and lock the door of the transmission window.

[0247] 2) Then transfer the sample to the second transmission window. Record the sample in the notebook and print 3 barcode labels. Then the sample is routinely tested and transferred to the pre-processing transmission window.

[0248] 2. Use a biological safety cabinet

[0249] 1) The hospital staff performing the pre-processing process enter the pre-processing studio and close the door. Open the bi...

Embodiment 3

[0277] Example 3. The process of using QIAamp Viral RNA kit to extract RNA

[0278] The following steps are used in the RNA preparation process:

[0279] 1. Use a pipette to add 560μl of the prepared buffer AVL containing Carrier RNA to a 1.5ml microcentrifuge tube. If the sample volume is greater than 140 μl, increase the amount of buffer AVL / Carrier RNA in the same proportion (for example, a 280 μl sample will require 1120 μl of buffer AVL / Carrier RNA).

[0280] 2. Add 140μl of plasma, serum, urine, cell culture supernatant or cell-free body fluid to the buffer AVL / Carrier RNA in the microcentrifuge tube. Mix for 15 seconds with a pulse vortex shaker. To ensure effective lysis, it is important to thoroughly mix the sample with buffer AVL to obtain a uniform solution. Frozen samples that have only been thawed once can also be used.

[0281] 3. Incubate for 10 minutes at room temperature (15-25°C). The virus particles can be completely lysed after 10 minutes at room temperature....

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Abstract

This invention relates generally to the field of virus detection. In particular, the invention provides chips, probes, primers, kits and methods for amplifying and detecting SARS-CoV nucleotides sequence. The clinical and other uses of the present chips, probes, primers, kits and methods are also contemplated.

Description

Background of the invention [0001] Since November 2002, a disease called Severe Acute Respiratory Syndrome (SARS) has been reported in 22 countries around the world. As of May 2, 2003, the WHO has reported a total of 6,054 SARS cases and 417 deaths among the infected people. At the same time, China reported a total of 3,788 SARS cases and 181 deaths among the infected population. [0002] The main symptoms of SARS patients include fever (above 38°C), headache and body aches. After the disease lasts for 2-7 days, the patient may develop a dry cough without sputum accompanied by breathing difficulties. [0003] Based on discoveries in Hong Kong, Canada and the United States, a previously unidentified coronavirus has been identified as the pathogen causing SARS. Researchers have discovered that SARS coronavirus is a positive-strand RNA virus that does not require intermediate steps in DNA during replication and uses standard codons (Marra et al., Science May 1, 2003; (before printing...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/70
CPCC12Q1/6837C12Q1/701C12Q1/686Y02A50/30
Inventor 李泽陶生策程京
Owner CAPITALBIO CORP
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