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Method of synchronous PCR detection of DNA and RNA pathogens with Tag enzyme

A pathogenic, 20% NP-40 technology, applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of wasting time and reagents, patients delaying treatment, etc., to save reagent costs, shorten detection time, The effect of reducing the chance of contamination

Inactive Publication Date: 2002-05-01
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

If primers and probes are used according to the existing data, it would be blind to carry out PCR amplification detection one by one.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0014] 1. Embodiment 1 Synchronous detection of HBV and HCV viral nucleic acid

[0015] Specific primers were designed according to the highly conserved regions of the published HBV and HCV genome sequences and submitted to Beijing Saibaisheng Company for synthesis. dNTP, TaqDNA polymerase, and 10×buffer (amplification buffer) were all purchased from Huamei Bioengineering Co., Ltd. Prepare our homemade protein lysis buffer.

[0016] The HBV upstream and downstream primers are: P1 5'-ACATACTCTGTGGAAGGCTGGC-3'P2 5'-TATCCCATGAAGTTAAGGGAGTAG-3'The length of the amplified fragment is about 430bp. HCV upstream and downstream primers are: P3 5'-TGCACGGTCTACGAGACCT-3'P4 5'-GCCATGGCGTTAGTATGAGT-3', the length of the amplified fragment is about 260bp.

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Abstract

A synchronous fast amplification PCR method for detecting DNA and RNA pathogens with Taq enzyme includes synchronously extracting DNA and RNA of virus nucleic acid by protein cracking reagent and boiling method, direct PCR amplification by using the polymerase activity and reverse transcription activity of Taq DNA polymerase and the virus nucleic acid as template to obtain desired nucleic acid fragments, and electrophoresis or hybridization for gene test. Its advantages are simplified process, low cost, short period, and less pollution chance.

Description

1. Technical field [0001] The invention relates to a gene detection technology, in particular to a method for rapidly amplifying and detecting DNA and RNA pathogens by simultaneous PCR (polymerase chain reaction) with Taq enzyme (heat-resistant DNA polymerase). 2. Background technology [0002] Gene research led to the rapid development of life science in the 20th century, and will continue to promote the in-depth development of life science research in the 21st century. Genetic testing is not only crucial to biological research, but also of great significance to fields such as clinical medicine, pharmacy, agricultural technology, environmental monitoring, and legal identification. [0003] The current PCR detection methods for DNA pathogens are mostly pre-treated samples, degraded protein shells, cell walls, cell membranes, etc. by alkali, surfactants, and proteases, and then extracted and separated waste proteins with phenol, chloroform-isoamyl alcohol, and then used Alco...

Claims

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Application Information

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IPC IPC(8): C12Q1/25C12Q1/68
Inventor 王业富庞代文沈钧涛
Owner WUHAN UNIV