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Real Time RT-PCR02:57

Real Time RT-PCR

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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...
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一步逆转录酶qPCR方法用于血清乙型肝炎病毒RNA量化

Shi Liu1, Bin Zhou1, Sheng Shen1

  • 1State Key Laboratory of Organ Failure Research, Guangdong Provincial Key Laboratory of Viral Hepatitis Research, Department of Infectious Diseases, Nanfang Hospital, Southern Medical University, Guangzhou, China.

Methods in molecular biology (Clifton, N.J.)
|July 23, 2024
PubMed
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此摘要是机器生成的。

血清乙型肝炎病毒 (HBV) RNA提供了一种非侵入性方法,用于评估慢性乙型肝炎 (CHB) 患者的肝内共闭环形DNA (cccDNA) 活性. 本协议详细说明了HBV RNA量化,以改善没有侵入性活检的临床管理.

关键词:
DNase I 在消化过程中.在HBV的RNA中,HBV的RNA是乙型肝炎病毒的感染.量化 量化 量化 量化逆转录酶 qPCRR 是一个

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科学领域:

  • 病毒学 病毒学
  • 肝病学 肝病学是一种肝病学.
  • 分子诊断学 分子诊断

背景情况:

  • 肝脏内共价封闭圆形DNA (cccDNA) 是乙型肝炎病毒 (HBV) 复制的模板,但其直接检测需要侵入性肝脏活检.
  • 慢性乙型肝炎 (CHB) 管理受到常规肝脏内ccDNA评估的不切实际性所阻碍.
  • 血清HBVRNA已经成为肝脏内ccDNA活性潜在的非侵入性生物标志物.

研究的目的:

  • 为量化血清HBVRNA提出一个详细的协议.
  • 为了确定血清HBVRNA作为反映肝脏内ccDNA活性的替代标记.
  • 提供一种非侵入性的替代方案,用于监测CHB患者的HBV复制.

主要方法:

  • 从患者血清样本同时分离HBV DNA和RNA.
  • DNase I消化以消除污染的HBV DNA.
  • 使用一步逆转录定量PCR (RT-qPCR) 试验量化HBV RNA.

主要成果:

  • 描述的协议使得可靠的血清HBVRNA的量化.
  • 血清HBVRNA水平与肝脏内ccDNA活性相关.
  • 该方法提供了一种非侵入性的方法来评估病毒活性.

结论:

  • 血清HBVRNA量化是一种可行的,非侵入性的方法来监测HBVccDNA活性.
  • 该协议可以通过提供对病毒复制的见解来帮助CHB患者的临床管理.
  • 这些发现支持使用血清HBVRNA作为B型肝炎研究和临床实践中有价值的生物标志物.