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PAXgene Blood RNA Tube (IVD)

For collection of whole blood samples for molecular diagnostic testing

  • Coupled with the PAXgene Blood RNA Kit (IVD), constitutes the FDA-cleared, CE-marked PAXgene Blood RNA System
  • Single tube for collection, transport, and storage
  • Immediate stabilization of intracellular RNA at collection
  • Specimen transport and storage at room, refrigerated or frozen temperatures
  • Frozen specimen storage for up to 8 years
  • Standardized preanalytic processing of samples

Feature

Specification

Tube size

16 x 100 mm

Blood draw volume

2.5 ml

Additive/concentration

Additive 6.9 ml

Length of stabilization

  • up to 3 days at room temperature (15–25°C)
  • up to 5 days at 2–8°C
  • 8 years at –20°C or –70°C*

Closure type/color

BD Hemogard/Red

Label type

Paper

Quantity

100/box

*Long-term study of blood storage in PAXgene Blood RNA Tubes is ongoing.


Intended Use

The PAXgene Blood RNA System consists of a blood collection tube (PAXgene Blood RNA Tube) and a nucleic acid purification kit (PAXgene Blood RNA Kit). It is intended for the collection, transport, and storage of blood and stabilization of intracellular RNA in a closed tube and subsequent isolation and purification of intracellular RNA from whole blood for RT-PCR used in molecular diagnostic testing.

Performance characteristics of the PAXgene Blood RNA System have been established with FOS and IL1B gene transcripts. The user is responsible for establishing appropriate PAXgene Blood RNA System performance characteristics for other target transcripts.
 

Order information

Product

Catalog Nr.

Price

PAXgene Blood RNA Tubes (100) IVD

100 blood collection tubes (2.5 ml). To be used in conjunction with PAXgene Blood RNA Kits.

762165
please inquire

Details

Performance

The patented additive in PAXgene Blood RNA Tubes protects RNA molecules from degradation by RNases and other enzymes, and triggers the complete lysis of all cells in the sample to minimize ex vivo changes in gene expression due to gene induction and downregulation. The result is immediate and long-term stabilization of RNA in specimens, even after being stored for up to 3 days at room temperature (Figure 1  and  Figure 2) or up to 5 days refrigerated (Figure 3  and Figure 4). Currently available data show stabilization of intracellular RNA for up to 8 years at –20°C or –70°C (see Technical Notes).

Principle

The collection of whole blood is the first step in many molecular assays used to study intracellular RNA. A major challenge in this type of testing is the instability of intracellular RNA, which rapidly degrades within hours after blood collection. Furthermore, certain transcript levels change after blood collection due to ex vivo gene regulation, such as induction and repression. Both RNA degradation and gene regulation can lead to an under- or overestimation of in vivo gene expression profiles.

The PAXgene Blood RNA Tube contains an additive that stabilizes the in vivo gene expression profile by reducing post-collection RNA degradation and minimizing gene induction and repression. When used in conjunction with one of the PAXgene Blood RNA Kits, the PAXgene Blood RNA Tube provides pure RNA for accurate detection and quantification of gene expression.

Procedure

Whole blood is collected directly into the PAXgene Blood RNA Tube, using standard phlebotomy practice and equipment. The RNA in the specimen is immediately stabilized for storage at room temperature, refrigerated, or frozen. RNA can be purified using one of the PAXgene Blood RNA Kits (choose the right PAXgene Blood RNA Kit). The ease of collection, stabilization of RNA at different temperatures, and elimination of additional processing steps (e.g., centrifugation, storage on ice) standardizes the workflow from collection to purification, reducing variability that can be introduced by different operators (Figure 5).

Applications

When used in conjunction with the PAXgene Blood RNA Kit, the PAXgene Blood RNA Tubes enable purification of intracellular RNA from whole blood for applications in molecular diagnostic testing, such as real-time RT-PCR analysis of specific transcripts.


PAXgene Blood RNA Tubes are intended for in vitro diagnostic testing (IVD) and are to be used in conjunction with the PAXgene Blood RNA Kit.

Blood was drawn from 10 donors, with duplicate samples, and stored at 18–25°C for the indicated number of days, followed by total RNA purification. [A] Blood was collected and stored in PAXgene Blood RNA Tubes, and total RNA was purified using the PAXgene Blood RNA Kit. [B] Blood was collected and stored in standard blood collection tubes with EDTA as an anticoagulant, and total RNA was purified using a standard organic-extraction method with silica-membrane–based RNA cleanup. Relative transcript levels of FOS were determined by real-time, duplex RT-PCR, using 18S rRNA as an internal standard. The values for all samples are plotted, with means and standard deviations of all samples shown. The dashed lines indicate the ±3x total precision of the assay (2.34 CT).Zoom image

Figure 1. RNA stability at 18–25°C: FOS.

Blood was drawn from 10 donors, with duplicate samples, and stored at 18–25°C for ...
Blood was drawn from 10 donors, with duplicate samples, and stored at 18–25°C for the indicated number of days, followed by total RNA purification. [A] Blood was collected and stored in PAXgene Blood RNA Tubes, and total RNA was purified using the PAXgene Blood RNA Kit. [B] Blood was collected and stored in standard blood collection tubes with EDTA as an anticoagulant, and total RNA was purified using a standard organic-extraction method with silica-membrane–based RNA cleanup. Relative transcript levels of IL1B were determined by real-time, duplex RT-PCR, using 18S rRNA as an internal standard. The values for all samples are plotted, with means and standard deviations of all samples shown. The dashed lines indicate the ±3x total precision of the assay (1.93 CT).Zoom image

Figure 2. RNA stability at 18–25°C: IL1B

Blood was drawn from 10 donors, with duplicate samples, and stored at 18–25°C for ...
Blood was drawn from 10 donors, with duplicate samples, and stored at 2–8°C for the indicated number of days, followed by total RNA purification. [A] Blood was collected and stored in PAXgene Blood RNA Tubes, and total RNA was purified using the PAXgene Blood RNA Kit. [B] Blood was collected and stored in standard blood collection tubes with EDTA as an anticoagulant, and total RNA was purified using a standard organic-extraction method with silica-membrane-based RNA cleanup. Relative transcript levels of FOS were determined by real-time, duplex RT-PCR, using 18S rRNA as an internal standard. The values for all samples are plotted, with means and standard deviations of all samples shown. The dashed lines indicate the ±3x total precision of the assay (2.34 CT).Zoom image

Figure 3. RNA stability at 2–8°C: FOS.

Blood was drawn from 10 donors, with duplicate samples, and stored at 2–8°C for ...
Blood was drawn from 10 donors, with duplicate samples, and stored at 2–8°C for the indicated number of days, followed by total RNA purification. [A] Blood was collected and stored in PAXgene Blood RNA Tubes, and total RNA was purified using the PAXgene Blood RNA Kit. [B] Blood was collected and stored in standard blood collection tubes with EDTA as an anticoagulant, and total RNA was purified using a standard organic-extraction method with silica-membrane-based RNA cleanup. Relative transcript levels of IL1B were determined by real-time, duplex RT-PCR, using 18S rRNA as an internal standard. The values for all samples are plotted, with means and standard deviations of all samples shown. The dashed lines indicate the ±3x total precision of the assay (1.93 CT).Zoom image

Figure 4. RNA stability at 2–8°C: IL1B.

Blood was drawn from 10 donors, with duplicate samples, and stored at 2–8°C for ...
Blood samples from 30 different donors were collected in PAXgene Blood RNA Tubes (12 tubes per donor, 360 tubes in total). The contents of the tubes from 3 donors were pooled and subsequently realiquoted into 36 samples. These 36 samples per 3-donor-pool were manually processed by 3 different operators. Each operator used 3 different PAXgene Blood RNA Kit lots for the extraction and processed quadruplicate samples from each of the 10 donor pools. Relative transcript levels of [A] FOS and [B] IL1B were determined by real-time, duplex RT-PCR using 18S rRNA as an internal standard. The values for all samples are plotted, relative to the values for user 1 (10 donor pools x 3 kit lots x 4 replicates = 120 data sets for each gene), with means (red lines) and standard deviations (black bars) for all samples shown. The dashed lines indicate the ±3x total precision of the assays (FOS, 2.34 CT; IL1B, 1.93 CT).Zoom image

Figure 5. Reproducibility between users.

Blood samples from 30 different donors were collected in PAXgene Blood RNA Tubes ...
Blood collected in duplicate from each of ten subjects was stored at –20°C. At each indicated time point, RNA was purified from duplicate tubes with the PAXgene Blood RNA Kit and analyzed using a quantitative RT-PCR assay specific for FOS. Mean ΔΔCT values for all time points are plotted as a red line with standard deviations. Red dashed lines represent assay precision (±3× total precision of the assay with consideration of single data; |3 × σT| = 1.16 CT).Zoom image

Figure 6. Relative transcript level changes of FOS in blood stored in situ at –20°C in PAXgene Blood RNA Tubes.

Blood collected in duplicate from each of ten subjects was stored at –20°C. At ...
Blood collected in duplicate from each of ten subjects was stored at –20°C. At each indicated time point, RNA was purified from duplicate tubes with the PAXgene Blood RNA Kit and analyzed using a quantitative RT-PCR assay specific for IL1B. Mean ΔΔCT values for all time points are plotted as a red line with standard deviations. Red dashed lines represent assay precision (±3× total precision of the assay with consideration of single data; |3 × σT| = 1.98 CT).Zoom image

Figure 7. Relative transcript level changes of IL1B in blood stored in situ at –20°C in PAXgene Blood RNA Tubes.

Blood collected in duplicate from each of ten subjects was stored at –20°C. At ...
Mean RNA integrity number (RIN) scores for duplicate specimens (subjects 1–10) and for all specimens in total (mean) are shown for the indicated storage times. The error bars indicate the upper and lower RIN scores of duplicate specimens from an individual subject or, for the mean column, the standard deviations of RIN scores of all specimens from all subjects.Zoom image

Figure 8. Integrity of RNA purified from blood stored in situ in PAXgene Blood RNA Tubes at –20°C.

Mean RNA integrity number (RIN) scores for duplicate specimens (subjects 1–10) and ...
Blood collected in duplicate from each of ten subjects was stored at –70°C. At each time point indicated, RNA was purified from duplicate tubes with the PAXgene Blood RNA Kit and analyzed using a quantitative RT-PCR assay specific for FOS. Mean ΔΔCT values for all time points are plotted as a red line with standard deviations. Red dashed lines represent assay precision (±3× total precision of the assay with consideration of single data; |3 × σT| = 1.16 CT).Zoom image

Figure 9. Relative transcript level changes of FOS in blood stored in situ at –70°C in PAXgene Blood RNA Tubes.

Blood collected in duplicate from each of ten subjects was stored at –70°C. At ...
Blood collected in duplicate from each of ten subjects was stored at –70°C. At each time point indicated, RNA was purified from duplicate tubes with the PAXgene Blood RNA Kit and analyzed using a quantitative RT-PCR assay specific for IL1B. Mean ΔΔCT values for all time points are plotted as a red line with standard deviations. Red dashed lines represent assay precision (±3× total precision of the assay with consideration of single data; |3 × σT| = 1.98 CT).Zoom image

Figure 10. Relative transcript level changes of IL1B in blood stored in situ at –70°C in PAXgene Blood RNA Tubes.

Blood collected in duplicate from each of ten subjects was stored at –70°C. At ...
Mean RNA integrity number (RIN) scores for duplicate specimens (subjects 1–10) and for all specimens in total (mean) are shown for the indicated storage times. The error bars indicate the upper and lower RIN scores of duplicate specimens from an individual subject or, for the mean column, the standard deviations of RIN scores of all specimens from all subjects.Zoom image

Figure 11. Integrity of RNA purified from blood stored in situ in PAXgene Blood RNA Tubes at –70°C.

Mean RNA integrity number (RIN) scores for duplicate specimens (subjects 1–10) and ...
Blood samples from 30 different donors were collected in PAXgene Blood RNA Tubes (12 tubes per donor, 360 tubes in total). The contents of the tubes from 3 donors were pooled and subsequently realiquoted into 36 samples. These 36 samples per 3-donor-pool were manually processed by 3 different operators. Each operator used 3 different PAXgene Blood RNA Kit lots for the extraction and processed quadruplicate samples from each of the 10 donor pools. Relative transcript levels of [A] FOS and [B] IL1B were determined by real-time, duplex RT-PCR using 18S rRNA as an internal standard. The values for all samples are plotted, relative to the values for kit lot 1 (10 donor pools x 3 users x 4 replicates = 120 data sets for each gene), with means (red lines) and standard deviations (black bars) for all samples shown. The dashed lines indicate the ±3x total precision of the assays (FOS, 2.34 CT; IL1B, 1.93 CT).Zoom image

Figure 12. Reproducibility between kit lots.

Blood samples from 30 different donors were collected in PAXgene Blood RNA Tubes ...
RNA was purified by 3 different operators (A, B, C) using 3 different lots (1, 2, 3) of the PAXgene Blood RNA Kit using the automated protocol in the experiment described in the figure "RNA yield and purity — automated processing". In parallel, RNA was purified from the corresponding replicate tubes using the manual protocol. Relative transcript levels of [A] FOS and [B] IL1B were determined by real-time, duplex RT-PCR using 18S rRNA as an internal standard. Possible differences of transcript levels between RNA prepared from paired blood samples using both extraction protocols (automated and manual protocol) were calculated by the ΔΔCT method. Individual ΔΔCT values for all sample pairs (4 replicates x 8 donor pools x 3 kit lots x 3 operators = 288 pairs for each gene) are plotted as single dots with means (larger dots) and standard deviations (black bars) for all samples shown. The dashed lines indicate the ±3x total precision of the assays (FOS: 2.34 CT; IL1B, 1.93 CT).Zoom image

Figure 13. Reproducibility between automated and manual protocols.

RNA was purified by 3 different operators (A, B, C) using 3 different lots (1, 2, ...
Blood samples from 48 different donors were collected in PAXgene Blood RNA Tubes (6 tubes per donor, 288 tubes in total). The contents of the tubes from 6 donors were pooled and subsequently realiquoted into 36 samples. These 36 samples per 6-donor-pool were processed by 3 different operators (A, B, C). Each operator used 3 different lots (1, 2, 3) of the PAXgene Blood RNA Kit for automated extraction and processed quadruplicate samples from each of the 8 donor pools. RNA yields, A260/A280 values, and genomic DNA amounts (w/w) of all individual samples are shown for every operator–lot combination.Zoom image

Figure 14. RNA yield, purity and genomic DNA contamination – automated processing.

Blood samples from 48 different donors were collected in PAXgene Blood RNA Tubes ...

Resources

PAXgene Blood RNA Tube

Phlebotomy and Sample Handling 

1. Why is a blood collection set required to be used with this tube?
A blood collection set is required to be used with the PAXgene Blood RNA Tube to eliminate the possibility of patient contact with the reagent in the tube, due to backflow of blood, when used in accordance with the instructions for use.

2. Is there any chance of the reagent going back into the patient's arm?
If used according to the instructions for use, there is no possibility of backflow during phlebotomy, and thus no possibility of reagent going into the patient’s arm.

3. Can another company's blood collection set be used with this tube?
Yes, as long as the phlebotomy instructions included with the PAXgene Blood RNA Tubes are properly followed.

4. In which order should the PAXgene Blood RNA Tube be drawn, if other tubes are used?
The PAXgene Blood RNA Tube should always be the last tube drawn.

5. Why do I have to use a discard tube?
The discard tube is used to fill the volume of the blood collection set tubing with blood so the subsequent draw into the PAXgene Blood RNA Tube is correct (2.5 ml). If other tubes are being drawn, they can also serve the purpose of a discard tube since the PAXgene Blood RNA Tubes are the last tubes drawn.

6. Can the PAXgene Blood RNA System be used with animal blood samples?
The PAXgene Blood RNA System was developed for intracellular RNA isolation from human blood samples. QIAGEN offers RNA stabilization and isolation systems for animal blood. Please contact QIAGEN technical service (www.qiagen.com) for more information.

7. Can the PAXgene Blood RNA System be used for human body fluids other than blood samples or cultured cells of human origin?
The PAXgene Blood RNA System was developed for intracellular RNA isolation from human blood specimens. PreAnalytiX offers a dedicated collection and RNA extraction system for human bone marrow (see PAXgene Bone Marrow RNA System). QIAGEN offers RNA stabilization and isolation systems for saliva and cultured cells. Please contact QIAGEN technical service (www.qiagen.com) for more information.

8. What are possible reasons for blood draws with lower than expected blood volume?
Possible reasons for blood draws with lower than expected blood volume include:
1) The phlebotomist has not waited enough time for the blood to stop flowing into the tube.
2) The tube has "pushed back" from the needle end attached to the holder and the tube is no longer drawing.
3) Discard tube was not used. For further information see the video Collecting Specimens In PAXgene Blood RNA Tubes and Instructions for Use.

9. Can blood be pipetted from an EDTA tube into the PAXgene Blood RNA Tube?
No. Transferring blood from any other tube into the PAXgene Blood RNA Tube is not recommended. The tube and its contents are optimized for performance on blood that is drawn directly into the tube at the recommended draw volume (2.5 ml). The use of anti-coagulated whole blood transferred by pouring or any other method is not recommended.

10. Can blood be injected from a syringe into the PAXgene Blood RNA Tube?
No. Using a syringe to collect and transfer blood into the PAXgene Blood RNA Tube is not recommended. This defeats the purpose of immediate stabilization of the sample that the direct-draw tube provides.

Transport and Storage

1. Can the PAXgene Blood RNA Tubes undergo freeze/thaw cycles?
PAXgene Blood RNA Tubes can go through up to two freeze/thaw cycles with no significant impact on yield or mRNA stability.

2. How should the PAXgene Blood RNA Tubes be frozen after collection?
The PAXgene Blood RNA Tubes should be frozen at –20°C and –70°C in a wire rack, not in an expanded polystyrene foam tray. If PAXgene Blood RNA Tubes are to be stored at temperatures lower than –20ºC, it is recommended to freeze them first for 24 h at –20ºC, and then transfer them to –70ºC.

3. How long can the tubes be kept frozen at –20°C or –70°C?
Currently we have data for storage at –20°C and –70°C for 96 months (In situ stability of RNA in blood specimens stored for 8 years (96 months) at –20°C and –70°C in PAXgene Blood RNA Tubes). Long term archiving studies are ongoing. See technical note: In Situ Stability of RNA in Blood Samples Stored at –20°C and –70°C in PAXgene Blood RNA Tubes.

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