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基于B型肝炎病毒的实时聚合酶连锁反应检测和定量DNA

Satyabrata Bag¹, Santanu Ghosh², Roopa Lalwani²

¹3B BlackBio Biotech India Limited; satya@3bblackbio.com.

Journal of visualized experiments : JoVE

|January 1, 2024

概括

本研究介绍了一种新的实时PCR套件,用于准确检测和定量乙型肝炎病毒 (HBV) DNA. 该测试为诊断和监测HBV感染提供了高灵敏度,有助于临床管理.

科学领域:

肝病学肝病学是一种肝病学.
分子诊断学分子诊断
病毒学病毒学

背景情况:

乙型肝炎病毒 (HBV) 感染是全球主要的健康问题,导致严重的肝脏疾病,如肝硬化和癌症.
准确量化HBVDNA对于诊断,监测治疗反应和管理HBV感染患者至关重要.

研究的目的:

用IVD标记的实时PCR套件在人体血清或血中检测和量化HBVDNA的详细方案.
评估套件在所有HBV基因型中的性能及其对临床诊断的敏感性.

主要方法:

使用IVD标记的实时PCR套件,针对HBV基因组的核心区域.
采用了针对所有HBV基因型 (A-J) 具有内源性内部控制的特定的原料和探针.
量化涉及一组5个标准与世卫组织第四个HBVDNA国际标准进行校准.

主要成果:

该试验显示出高灵敏度,检测HBV DNA的低至2.5 IU/mL.
实时PCR协议准确量化了所有测试的基因型中的HBVDNA.
在分析过程中,内部控制有效地监测了潜在的PCR抑制.

结论:

开发的实时PCR套件为HBV DNA检测和定量提供了一种敏感和可重复的方法.
这种工具对临床实验室在有效诊断和治疗乙型肝炎病毒感染方面非常有价值.

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Real Time RT-PCR

Real Time RT-PCR

Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...