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相关概念视频

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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Simultaneous Quantification of T-Cell Receptor Excision Circles TRECs and K-Deleting Recombination Excision Circles KRECs by Real-time PCR
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在常规测试中实现实时PCR结果的规范化.

Betsy Armenta-Leyva1, Berenice Munguía-Ramírez1, Ting-Yu Cheng2

  • 1Department of Veterinary Diagnostic and Production Animal Medicine, College of Veterinary Medicine, Lloyd Veterinary Medical Center, Iowa State University, Ames, IA, USA.

Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc
|November 3, 2023
PubMed
概括

这项研究引入了效率标准化的Cqs (ECqs) 来实现实时PCR测定的正常化,改善了在血清和口服液样本中检测猪生殖和呼吸系统综合征病毒 (PRRSV) 的检测. 该ECq方法提高了PRRSV监测的诊断准确性.

关键词:
ECq ECq ECq ECq ECq ECq ECq ECq ECq ECq ECq ECq放大功率的效率是放大功率的效率.规范化的正常化.在实时PCR中进行实时PCR.运行到运行变化的变化.

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科学领域:

  • 兽医诊断 兽医诊断 兽医诊断 兽医诊断
  • 分子生物学分子生物学
  • 病毒学 病毒学

背景情况:

  • 实时PCR测定需要正常化,以控制采样和测试的变化.
  • 实时PCR中的量化周期 (Cqs) 可以转换为效率标准化的Cqs (ECqs),以提高准确性.
  • 猪生殖和呼吸系统综合征病毒 (PRRSV) 对猪健康和经济构成重大威胁.

研究的目的:

  • 开发和验证一个ECq方法来规范商业PRRSVRT-qPCR测定.
  • 评估ECq方法在血清和口服液样本中的诊断性能.
  • 提高在猪中检测PRRSV的准确性和可靠性.

主要方法:

  • 使用公式E-ΔCq计算ECqs,其中E是放大效率,ΔCq是样本和参考标准Cqs之间的差异.
  • 使用在血清或口服液中稀释的PRRSV改性活疫苗创建了参考标准.
  • 来自接种疫苗的猪的血清和口液样本进行了测试,Cqs被转换为ECqs.

主要成果:

  • 该ECq方法显示出高的诊断准确性,血清的曲线下的面积为0.999和口服液的0.947.
  • 两种样本类型的诊断灵敏度和特异性都很高,血清显示97.9%的灵敏度和100%的特异性,口服液显示82.6%的灵敏度和100%的特异性.
  • 放大效率 (E) 范围为血清1.7~2.6,口服液1.7~2.3.

结论:

  • 该ECq方法提供了一个强大的方法来规范PRRSVRT-qPCR测定.
  • 这种规范化策略提高了在血清和口腔液体中检测PRRSV的诊断灵敏度和特异性.
  • 该ECq方法有望改善PRRSV监测和猪群疾病管理.