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Diagnosis reagent box for detecting SARS coronavirus using transcription PCR method

A diagnostic kit and coronavirus technology, applied in biochemical equipment and methods, microbiological determination/inspection, resistance to vector-borne diseases, etc., can solve the problems of no clinical statistical data publication, different sequences, no comparability, etc. , to achieve the effect of increased sensitivity, high sensitivity and high accuracy

Inactive Publication Date: 2004-07-21
珍奥集团股份有限公司
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Problems solved by technology

[0026] It can be seen that the primers used in the design of each RT-PCR are different, and the sequences compared are also different; so it is difficult to say which one is more effective, and, so far, there is no clinical statistical data published; if only In terms of efficiency, it is meaningless and not comparable, which is why WHO calls for the development of more accurate PCR detection methods

Method used

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  • Diagnosis reagent box for detecting SARS coronavirus using transcription PCR method
  • Diagnosis reagent box for detecting SARS coronavirus using transcription PCR method
  • Diagnosis reagent box for detecting SARS coronavirus using transcription PCR method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0108] Example 1. Kit 1

[0109] 1) Materials: Positive specimens are from the National Academy of Military Medical Sciences, and the samples are genetically engineered Vero-6 virus cell strains with SARS virus RNA; stored at -80C for future use. Control substances were normal human saliva and other viruses.

[0110] 2) Preparation of RNA samples: the preparation of RNA samples refers to NEJM (document 13.Marco A.Marra, Steven J.M.Jones, Caroline R.Astell etc, The Genome Sequenceof the SARS-Associated Coronavirus, Science, www.scienceexpress.org1 May 2003 ), that is, add an equal volume of N-acetyl cysteine ​​buffer solution (0.9% NaCl fusion solution, the concentration is 10g / l N-acetyl cysteine ​​buffer solution) in the body fluid containing SARS cells; slowly Shake for 30 minutes; take 600 μL of this solution, centrifuge (10,000 g) in a conventional tabletop centrifuge (10,000 g) for 3 minutes, add 140 μL of supernatant to 560 μL of AVL buffer (Qiagen viral RNA kit), and r...

Embodiment 2

[0226] Embodiment 2. Kit 2

[0227] Primers used primer pair BIT001, and other parts of the kit were the same as Example 1.

Embodiment 3

[0228] Embodiment 3. Kit 3

[0229] Primers used primer pair BIT002, and other parts of the kit were the same as Example 1.

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Abstract

A reagent kit for detecting SARS coronavirus by reverse transcription PCR method has buffering liquid system and the cDNA fragment as template relative to the Orflb reverse transcription in RNA of SARS coronavirus. Its primer pair is BIT 001-009. Its advantages are high sensitivity, correctness, specificity and speed, and low cost.

Description

technical field [0001] The present invention relates to the diagnosis of SARS coronavirus, specifically a kind of reverse transcription PCR method detects SARS coronavirus diagnostic kit. Background technique [0002] Severe Acute Respiratory Syndrome (Severe Acute Respiratory Syndrom, SARS), or Atypical Pneumonia (Atypical Pneumonia); it began to occur in Guangdong around November 2002, and then spread rapidly to all parts of the world, causing a global outbreak in the spring of 2003 Disease; its rapid onset, deep harm, and wide impact are the most serious ones since this century; for this reason, the World Health Organization (WHO) issued a global early warning on March 12 this year, and united 10 countries around the world A cooperative research network composed of 11 top laboratories in China and the region (Document 1. WHO WHO issues a global alert about cases of atypical pneumonia. March 12, 2003. Document 2. WHO China joins WHO collaborative network. March 28, 2003), ...

Claims

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Application Information

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IPC IPC(8): C12Q1/04C12Q1/68
CPCY02A50/30
Inventor 梅晓丹周小棉刘大渔周润涛夏志南张利平林炳承
Owner 珍奥集团股份有限公司
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