Yields of Viral and Circulating Cell-Free Nucleic Acids Using the QIAamp® Circulating Nucleic Acid Kit

Abstract

Fragmented DNA and RNA circulate as cell-free nucleic acids in plasma, serum, urine and other body fluids. Access to these molecules for analysis may allow for detection of certain disease states based on a blood sample. In this study the extraction efficiency of a large volume nucleic acid extraction kit for circulating and viral nucleic acids was assayed with different downstream qPCR assays. For circulating DNA in plasma the yield was 7-to 9-fold higher with the large volume kit compared to the QIAamp DNA Blood Mini protocol with improved recovery of short DNA fragments. The microRNA extraction protocol of the large volume kit was found to deliver significantly increased microRNA yields ( Ct from 3 to 4) compared to the kit's standard protocol. For viral nucleic acids, 95% detection rates were: HBV DNA 0.25 IU/ml and HIV-1 RNA 14.2 IU/ml. The results demonstrate that the QIAamp Circulating Nucleic Acid Kit can serve as a sample preparation solution for processing up to 5 ml cell-free body fluid, it allows for small elution volumes, and can extract and concentrate circulating nucleic acids, including microRNA, and viral nucleic acids up to 250 fold.

Circulating nucleic acids in plasma and serum: diagnosis and prognosis in cancer

“…A recent development of the QIAamp circulating nucleic acid kit working with larger volumes of either plasma or serum may offer a more complete extraction kit [ 27 ].…”

Benchmarking blood collection tubes and processing intervals for extracellular vesicle performance metrics

“…In the EV Blood Benchmarking (EVBB) study, we evaluated the impact of 11 BCT, including serum, four routinely used anticoagulants in clinical settings (non‐preservation BCT) and six specialized preservation additives (preservation BCT), on nine performance metrics including blood sample quality, EV stability, EV blood cell origins and EV‐associated proteome and small RNA transcriptome signatures (Figure 1). An overview of the selected BCT and their intended use, composition, mode of action and predicted effect on blood cells is provided in Table 1 (Banfi et al., 2007; Das et al., 2017; Fernando, 2017; Fernando & Chao‐Wei, 2016; Horlitz et al., 2020; Mannuß, 2020; Muller et al., 2018). Since sample processing within a short timeframe may be difficult to achieve in a clinical setting, five BCT with favourable performance metrics for EV analysis were additionally evaluated for three clinically relevant sample BPI (Figure 1).…”

Reliable Determination of Fetal RhD Status by <b><i>RHD </i></b>Genotyping from Maternal Plasma

“…Therefore automated methods are preferable. There are some recommended manual methods such as the QIAamp DSP Virus Kit (Qiagen, Hilden, Germany) when compared to the QIAamp Blood Mini Kit (Qiagen) [15,24] or the recently released commercial QIAamp Circulating Nucleic Acid Kit (Qiagen) [25]. The recommended automated methods for the extraction of cff-DNA are the MagnaPure LC System (Roche Diagnostics, Basel, Switzerland) and the Magnetic Separation Module 1 (Chemagen, Baesweiler, Germany) [26], which allow high-throughput isolation.…”