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EZ1 DSP Virus Kit

应用EZ1 Advanced或EZ1 Advanced XL从1–6个或1–14个人类血浆、血清或脑脊液样本中自动、同步地纯化病毒DNA和RNA

Products

EZ1 DSP Virus Kit适用于体外诊断。
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EZ1 DSP Virus Kit (48)

Cat. No. / ID:   62724

For 48 viral nucleic acid preps: Prefilled Reagent Cartridges, Disposable Tip-Holders, Disposable Filter-Tips, Sample Tubes, Elution Tubes, buffers, carrier RNA
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特点

  • 更高的分析灵敏度，可用于处理低滴度病毒
  • 每轮可快速、灵活处理1–6个或1–14个样本
  • 程序设置简单，只需插卡即可运行
  • 标准化的流程，确保可重复性的结果

产品详情

EZ1 DSP Virus Kit包含所有需要的试剂和实验耗材，可运用磁性颗粒技术从人体血清、血浆、脑脊髓液（CSF）、尿液、全血、粪便、传输介质、呼吸样本和干拭子中自动纯化病毒核酸和细菌DNA。试剂都以预装试剂盒的形式提供，确保配合EZ1 Advanced或EZ1 Advanced XL使用的速度和便利性。EZ1 DSP Virus Kit具有精确的性能规格，确保高度可靠的病毒核酸纯化。

绩效

检测阈限低，分析灵敏度高

EZ1 DSP Virus Kit能够从人类血清、血浆、脑脊液、尿液、全血、粪便、呼吸道样本、干燥拭子中同时纯化病毒核酸和细菌DNA。使用CE认证的artus PCR Kits在多种real-time PCR仪上检测CMV DNA和HBV DNA，结果显示高效纯化获得的病毒RNA和DNA在PCR反应中表现出高分析灵敏度（参见下表）。

使用EZ1 DSP Virus检测产品和artus CMV PCR Kits对CMV DNA的检测阈限
病毒滴度（copies/ml） 检测阳性次数（LightCycler） 检测阳性次数（Rotor-Gene） 检测阳性次数（ABI PRISM）
1000 18/18 18/18 6/6
316 15/15 15/15 18/18
100 23/24 24/24 18/18
31.6 30/33 27/27 18/18
15.8 10/18 11/12 18/18
10 12/36 21/36 34/36
7.9 8/18 7/18 15/18
5.0 4/18 11/18 13/18
3.16 2/18 6/18 26/36
1 0/18 1/18 4/18
0.316 1/18 0/18 1/18
0.1 1/18 1/18 0/18
0 0/6 0/6 0/18
95%概率值(copies/ml) 67.2 21.8 13.2
置信区间(copies/ml) 41.8–142 14.5–44.1 9.0–23.1
The detection limit was determined by the 95 % probit value for the EZ1 DSP Virus system using quantified CMV cell-culture supernatant. The detection limit was determined by processing a dilution series of CMV. The virus was diluted in CMV-negative normal human EDTA plasma pool. Each dilution step was prepared in at least 3 independent runs with at least 6 replicates per dilution. Plasma (400 μl) was used for sample preparation with elution in 60 μl. artus CMV PCR Kits were used for detection of CMV DNA. The samples were analyzed on a LightCycler 1.2 Instrument (Roche), a Rotor-Gene 3000 (Corbett-Research), and an ABI PRISM 7000 SDS (Applied Biosystems).
使用EZ1 DSP Virus检测产品和artus HBV PCR Kits对HBV DNA的检测阈限
病毒滴度（copies/ml） 检测阳性次数（LightCycler） 检测阳性次数（Rotor-Gene） 检测阳性次数（ABI PRISM）
1000 6/6 12/12 18/18
316 18/18 18/18 15/15
100 18/18 18/18 24/24
31.6 18/18 18/18 32/33
15.8 10/18 18/18 16/18
10 13/36 34/36 26/36
7.9 9/18 15/17 13/18
5.0 4/18 13/18 9/18
3.16 5/36 23/36 7/18
1 1/18 5/15 5/18
0.316 0/18 2/17 2/18
0.1 0/18 1/18 2/18
0 0/18 0/18 0/6
95%概率值(copies/ml) 45.7 14.4 38.3
置信区间(copies/ml) 28–102 9.5–26.5 21.5–89.8
The detection limit was determined by the 95% probit value for the EZ1 DSP Virus system using HBV WHO international virus standard. The detection limit was determined by processing a dilution series of HBV. The virus was diluted in HBV-negative normal human EDTA plasma pool. Each dilution step was prepared in at least 3 independent runs with at least 6 replicates per dilution. Plasma (400 μl) was used for sample preparation with elution in 60 μl. artus HBV PCR Kits were used for detection of HBV DNA. The samples were analyzed on a LightCycler 1.2 Instrument (Roche), a Rotor-Gene 3000 (Corbett-Research), and an ABI PRISM 7000 SDS (Applied Biosystems).
高纯度病毒DNA和RNA产量呈线性

病毒核酸的纯化产量呈线性，因此无论高还是低病毒滴度，都能获得准确的定量分析结果。纯化的病毒核酸在敏感的分析中具有出色表现，即便起始样本中存在高水平内源性抑制剂也能实现灵敏检测（参见下表）。

可靠的纯化，去除抑制剂
抑制剂 浓度 平均log (copies/ml) 标准差
未处理 – 2.87 0.04
胆红素 20 mg/dl 2.83 0.02
血红蛋白 500 mg/dl 2.80 0.09
蛋白质 9 g/dl 2.77 0.06
Liposyn 3000 mg/dl 2.80 0.06
HBV (1000 copies/ml [log 3]) was added to panels of recalcified plasma containing elevated levels of endogenous inhibitors. A 3.4 μl aliquot of RealTime HBV Internal Control was combined with carrier RNA for each sample, and viral nucleic acids were extracted from 400 μl samples, in replicates of 5, and eluted in 90 μl elution buffer (AVE). The Abbott RealTime HBV assay was used for detection of HBV. PCR was carried out on the Abbott m2000rt.
无样本交叉污染

核酸纯化的过程可靠性对实验室常规检测极为重要。样本批次间无交叉污染是获得可靠结果的先决条件。为评估不同批次间的样本交叉污染，使用阴性阳性（1.0 x 108 IU/ml）B19微小病毒样本间隔排布的孔板严格检测EZ1 DSP Virus检测产品的性能。使用CE认证的artus Parvo B19 RG PCR Kit成功检测出所有高度阳性样本。所有阴性样本未检出（参见下表）。这表明，EZ1 DSP Virus检测产品在上述实验条件下无样本交叉污染。

交叉污染检测，检测B19微小病毒DNA的检测CT值
孔板位置
批次 1 2 3 4 5 6
1 15.47 X 15.41 X 15.36 X
2 X 15.48 X 15.53 X 15.32
3 X X X X X X
4 15.35 X 15.20 X 15.27 X
5 X 15.21 X 15.13 X 15.42
6 X X X X X X
7 15.X2 X 15.48 X 15.23 X
8 X 15.31 X 15.83 X 15.62
9 X X X X X X
Nine runs were performed to evaluate the risk of cross contamination events during and between EZ1 DSP Virus procedures. The test was performed using a quantified parvovirus B19 patient sample. The viral load of positive samples used for the carryover tests was 1.0 x 108 IU/ml. For dilution of positive samples and as negative control samples, a human parvovirus B19 negative EDTA plasma pool was used. To detect sample-to-sample carryover, 2 runs were performed with an alternating checkerboard setup of negative and highly positive samples. Every third run was performed using all negative samples to monitor possible run-to-run carryover. This sample setup was repeated three times resulting in a total of nine runs. Parvovirus B19 DNA was detected and quantitated using the CE-IVD-marked artus Parvo B19 RG PCR Kit on the Rotor-Gene 3000. The analytical detection limit of the artus Parvo B19 RG PCR Kit is determined to be 0.2 IU/μl in the eluate (p = 0.05). This indicates that there is a 95 % probability that 0.2 IU/μl in the eluate will be detected. Mean CT value of all samples = 15.40 ± 0.18 (CV = 1.14%) X: Unresponsive after 45 PCR cycles.

原理

自动化样本制备确保高效、标准化的核酸纯化。EZ1 Advanced和EZ1 Advanced XL工作站使用经验证的磁珠技术纯化核酸。试剂产品受严格的质量控制，确保可靠、稳定的性能。易用的仪器、预封装的试剂条、简单的实验方案和操作平台的设置有助于减少手动操作的误差，无论是初学者还是熟练者均可运行该体系。尽量减少了手动操作可能具传染性样本，确保使用者的安全和可靠的样品处理过程。纯化的病毒DNA和RNA可即用于基于酶扩增的敏感下游诊断分析，如通过PCR和反转录PCR（RT-PCR）进行病毒量监测。

程序

EZ1 Advanced或EZ1 Advanced XL工作站与经验证的EZ1 DSP Virus Kit配合使用，可从1–6（EZ1 Advanced）或1–14（EZ1 Advanced XL）个样本中纯化病毒核酸和细菌DNA，样本包括人体血清、血浆、CFS、尿液、全血、粪便、传输介质、呼吸样本和干拭子。从穿透试剂条到洗脱高纯度核酸，所有操作流程由工作站执行。该工作站的创新点在于将吸量管作为分离腔、提高磁珠纯化的效率和避免一系列离心操作。优化的操作步骤使手动操作时间最少，实现高度自动化的核酸纯化。

应用

EZ1 DSP Virus Kit配合EZ1 Advanced DSP Virus Card和EZ1 Advanced或者EZ1 Advanced XL DSP Virus Card和EZ1 Advanced XL，提供了从人体血清、血浆、CSF、尿液、全血、粪便、传输介质、呼吸样本和干拭子中同时纯化病毒核酸和细菌DNA的全自动操作流程。使用EZ1 DSP Virus Kit纯化的病毒核酸可即用于基于PCR和RT-PCR等酶扩增反应的下游诊断分析。

Specifications

FeaturesSpecifications
For automated processingBioRobot EZ1
ApplicationsZHPCR, real-time RT-PCR, genotyping
ProcessingAutomated
Purification of total RNA, miRNA, poly A+ mRNA, DNA or proteinViral DNA, viral RNA
TechnologyMagnetic particles
CE/FDA/IVD compatibleCE/IVD
Main sample typeSerum, plasma
Sample amount100–400 µl
Elution volume75–50 µl

资源

应用说明 (6)
Automated purification of DNA from buffy coat samples using the BioRobot EZ1 System - (EN)
PDF (480KB)
(EN) - Routine use of the BioRobot EZ1 in detection of veterinary and zoonotic pathogens
PDF (448KB)
Automated DNA preps from soft tissue biopsies and mouse tails using the BioRobot EZ1 System - (EN)
PDF (484KB)
High-performance DNA from forensic samples using the EZ1 DNA Tissue Kit together with the EZ1 DNA Forensic Card protocols - (EN)
PDF (411KB)
(EN) - Reproducible purification of DNA from blood using the BioRobot EZ1 System
PDF (500KB)
(EN) - Efficient purification of high-quality DNA from blood samples with a range of white cell counts using the BioRobot EZ1 System
PDF (477KB)
试剂盒操作手册 (2)
(EN) - EZ1 DSP Virus Kit Handbook (Version 4)
PDF (1MB)
Version 4
EZ1 DSP Important Note
PDF (793KB)
New 2D barcodes on EZ1 DSP reagent cartridges
安全数据表 (1)
Safety Data Sheets
Download Safety Data Sheets for QIAGEN product components.
Certificates of Analysis (1)
Certificates of Analysis

FAQ

How long can eluates be stored after sample preparation using the BioRobot EZ1 DSP instrument?
Eluates can be stored for up to 10 weeks at 2-8°C; up to one year either at -20°C or at -80°C without any loss of performance.
FAQ ID -1569
What do I have to do if precipitates are visible in the lysis buffer (well 1 of the reagent cartridge [RCB])?
Place the reagent cartridges (RCB) into a shaker–incubator, and incubate at 30–40°C with mild agitation for up to 2 hours. Do not use the reagent cartridges (RCB) if the precipitates do not redissolve.
FAQ ID -1568
Can I use sample preparation cartridges from other suppliers together with the BioRobot EZ1 DSP instrument?
No. The BioRobot EZ1 DSP instrument is intended to be used only with QIAGEN sample preparation cartridges.
FAQ ID -1567
How long can eluates be stored after sample preparation using the EZ1 instruments or EZ2 Connect MDx and the EZ1 DSP Virus Kit?
For short-term storage of up to 24 hours, it is recommended to store the purified viral nucleic acids or bacterial DNA at 2–8°C. For long-term storage of more than 24 hours, it is recommended to store at –80°C for up to 12 months or –20°C for up to 12 weeks. Stability of nucleic acids might be different for the specific downstream application being used and needs to be self-validated by the user.
FAQ ID - 3897
How can I track samples during collection, sample preparation, and downstream assay?
A sample sheet template is included in the handbook. This sample sheet may be useful for recordkeeping when using the EZ1 DSP DNA Blood or the EZ1 DSP Virus procedure. This sheet can be photocopied and labeled with descriptions of the samples and details of the run.
FAQ ID -1570
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