InVita Healthcare Technologies, Jacksonville Beach, FL, USA.
Methods Mol Biol. 2023;2685:189-204. doi: 10.1007/978-1-0716-3295-6_12.
The QIAGEN Investigator Quantiplex Pro Kit is a real-time quantitative PCR assay utilized by forensic DNA laboratories to determine the amount of amplifiable human and male DNA in a sample prior to downstream amplification of specific STR markers for human identity testing. This quantification method includes two internal controls that assist the analyst in a preliminary evaluation of the sample in regard to both inhibition or degradation that may be present in the sample and subsequently affect the more targeted downstream amplification of specific markers for identity testing. The internal controls are analogous to the quality sensors contained in QIAGEN's Investigator 24plex line of amplification kits, ensuring that the sample's performance in the quantitation step can accurately predict the success of the STR amplification results. This chapter describes the physical plate setup of a quantitative PCR assay utilizing the QIAGEN Investigator Quantiplex Pro Kit as well as the steps and software configurations involved in running such a plate on the Applied Biosystems 7500 Real-Time PCR System for Human Identification using HID Real-Time PCR Analysis Software v1.1 or 1.2.
QIAGEN Investigator Quantiplex Pro 试剂盒是一种实时定量 PCR 检测试剂盒，被法医 DNA 实验室用于在下游扩增特定 STR 标记物进行人类身份检测之前，确定样本中可扩增的人类和男性 DNA 的含量。该定量方法包括两个内部对照物，可帮助分析人员初步评估样本中是否存在抑制或降解等情况，这些情况可能会影响后续针对特定标记物的更有针对性的下游扩增。内部对照物类似于 QIAGEN 的 Investigator 24plex 扩增试剂盒中包含的质量传感器，确保定量步骤中样本的性能能够准确预测 STR 扩增结果的成功。本章描述了使用 QIAGEN Investigator Quantiplex Pro 试剂盒进行定量 PCR 检测的物理板设置，以及在 Applied Biosystems 7500 Real-Time PCR 系统上运行此类板的步骤和软件配置，该系统使用 HID Real-Time PCR Analysis Software v1.1 或 1.2 进行人类识别。
