Highly sensitive and specific fluorescent quantitative PCR (Polymerase Chain Reaction) detection system and detection method for HBV (Hepatitis B Virus) pgRNA (pregenome Ribonucleic Acid) in blood

A fluorescence quantification, blood technology, applied in DNA/RNA fragments, biochemical equipment and methods, microbial determination/inspection, etc., can solve problems such as sensitivity and specificity need to be improved

Active Publication Date: 2017-08-18
PEKING UNIV
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Problems solved by technology

However, the sensitivity and specificity of the current fluorescent quantitative PCR method for detecting blood HBV RNA need to be improved

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  • Highly sensitive and specific fluorescent quantitative PCR (Polymerase Chain Reaction) detection system and detection method for HBV (Hepatitis B Virus) pgRNA (pregenome Ribonucleic Acid) in blood
  • Highly sensitive and specific fluorescent quantitative PCR (Polymerase Chain Reaction) detection system and detection method for HBV (Hepatitis B Virus) pgRNA (pregenome Ribonucleic Acid) in blood
  • Highly sensitive and specific fluorescent quantitative PCR (Polymerase Chain Reaction) detection system and detection method for HBV (Hepatitis B Virus) pgRNA (pregenome Ribonucleic Acid) in blood

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[0026] Unless otherwise specified, terms in the present invention have meanings commonly used in the art.

[0027]The term "nucleotide" is intended to include those moieties that contain not only the known purine and pyrimidine bases but also modified other heterocyclic bases. Such modifications include methylated purines or pyrimidines, acylated purines or pyrimidines, alkylated ribose sugars or other heterocyclic compounds. Furthermore, the term "nucleotide" includes those moieties which contain haptens or fluorescent labels and which may contain not only conventional ribose and deoxyribose sugars but also other sugars. Modified nucleosides or nucleotides may also comprise modifications on the sugar moiety, for example, wherein one or more hydroxyl groups are replaced with halogen atoms or aliphatic groups, functionalized as ethers, amines, and the like.

[0028] The term "primer" refers to a short, chemically synthesized oligonucleotide, typically about 20 to 30 bases in l...

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Abstract

The invention discloses a fluorescent quantitative PCR (Polymerase Chain Reaction) detection system for HBV (Hepatitis B Virus) pgRNA (pregenome Ribonucleic Acid) in blood and a method for detecting the HBV pgRNA by utilizing the detection system. The detection system comprises a forward primer, a reverse primer, a reverse transcription primer with a random anchoring primer and a probe, wherein the nucleotide sequence of the forward primer is as shown in SEQ NO: 1; the nucleotide sequence of the reverse primer is as shown in SEQ NO: 2; the nucleotide sequence of the reverse transcription primer is as shown in SEQ NO: 3; the nucleotide sequence of the probe is as shown in SEQ NO: 4. The detection system and the method which are provided by the invention are adopted to be capable of being used for preparing an HBV diagnostic reagent or kit. By using the detection system and the method, the possible interference brought by HBV DNA (Deoxyribonucleic Acid) or other RNA in a detection process is eliminated; the detection specificity of the HBV pgRNA in the blood is ensured.

Description

technical field [0001] The present invention generally relates to the diagnosis and treatment of viral hepatitis B, as well as the fields of infectious disease, molecular biology, cell biology and the like. More specifically, the present invention relates to a fluorescent quantitative PCR detection method for hepatitis B virus (Hepatitis B virus, HBV) pgRNA in blood. Background technique [0002] Hepatitis B virus (HBV) infection is prevalent in the world, but the epidemic intensity of HBV infection varies greatly in different regions. my country belongs to the high prevalence area of ​​HBV infection. The national HBV blood epidemiological survey in 2006 showed that the HBsAg carrier rate among the general population aged 1-59 in my country was 7.18%. Based on this calculation, there are about 93 million chronic HBV-infected people in my country, including about 20 million chronic hepatitis B (chronic hepatitis B) patients. In my country, chronic HBV infection is closely ...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11
CPCC12Q1/6851C12Q1/706C12Q2531/113C12Q2563/107
Inventor 鲁凤民王杰陈然
Owner PEKING UNIV
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