高灵敏度检测病毒RNA或DNA
  • 高灵敏度的单一和多重检测
  • 在同一反应中检测病毒RNA和/或DNA
  • 清晰检测弱阳性信号
  • 通用的快速两步法实验方案
  • 5x预混液灵敏度更高,起始样本量更大
QuantiTect Virus Kit专用于病毒核酸的高灵敏检测,使用序列特异性的探针,通过多重一步法real-time RT-PCR可对多至4个病毒核酸靶标进行检测。多重分析可对含有内参的多种病毒RNA和/或DNA靶分子进行检测,不会影响灵敏度。试剂盒中浓缩的5x预混液可实现更大上样量,还包含HotStarTaq Plus DNA Polymerase。QuantiTect Virus RT Mix含有独特设计的Sensiscript Reverse Transcriptase,经过优化可进行病毒RNA的高灵敏度检测。该试剂盒有两种规格:QuantiTect Virus Kit适用于需要ROX染料进行荧光校正的PCR仪,QuantiTect Virus +ROX Vial Kit适用于其他PCR仪。为便于使用,预混液可储存在2–8°C。
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QuantiTect Virus Kit (50)
Trial kit for 50 x 50 µl reactions: QuantiTect Virus Master Mix (contains ROX dye), QuantiTect Virus RT Mix, RNase-Free Water, QuantiTect Nucleic Acid Dilution Buffer
211011
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QuantiTect Virus Kit (200)
For 200 x 50 µl reactions: QuantiTect Virus Master Mix (contains ROX dye), QuantiTect Virus RT Mix, RNase-Free Water, QuantiTect Nucleic Acid Dilution Buffer
211013
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QuantiTect Virus Kit (1000)
For 1000 x 50 µl reactions: QuantiTect Virus Master Mix (contains ROX dye), QuantiTect Virus RT Mix, RNase-Free Water, QuantiTect Nucleic Acid Dilution Buffer
211015
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QuantiTect Virus +ROX Vial Kit (50)
Trial kit for 50 x 50 µl reactions: QuantiTect Virus NR Master Mix (without ROX dye), ROX Dye Solution, QuantiTect Virus RT Mix, RNase-Free Water, QuantiTect Nucleic Acid Dilution Buffer
211031
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QuantiTect Virus +ROX Vial Kit (200)
For 200 x 50 µl reactions: QuantiTect Virus NR Master Mix (without ROX dye), ROX Dye Solution, QuantiTect Virus RT Mix, RNase-Free Water, QuantiTect Nucleic Acid Dilution Buffer
211033
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QuantiTect Virus +ROX Vial Kit (1000)
For 1000 x 50 µl reactions: QuantiTect Virus NR Master Mix (without ROX dye), ROX Dye Solution, QuantiTect Virus RT Mix, RNase-Free Water, QuantiTect Nucleic Acid Dilution Buffer
211035
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QuantiTect Virus Kits are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.


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Unambiguous determination of CT values over a wide dynamic range.|Reliable detection of viral RNA over a wide linear range.|QIAGEN multiplex kits.|Unique PCR buffer.|Improved detection of low amounts of viral RNA.|Reliable dilution and storage of RNA standards.|
Serial dilutions of bovine viral diarrhea virus (BVDV) RNA, as indicated, were amplified and analyzed using the QuantiTect Virus Kit. The steep sigmoidal curves enable accurate CT value determination, even at low template amounts (10 copies/μl). [A] Detection of BVDV1 RNA. [B] Detection of BVDV2 RNA. NTC: No template control.|Viral RNA was diluted in serial fivefold dilutions and amplified in duplex with an internal control using the QuantiTect Virus Kit. [A] Amplification plot of the viral targets. [B] Amplification plot of the internal control.|The QuantiTect Virus Kit provides a simple procedure for quantitative, multiplex, real-time PCR. In contrast to current methods, the kits eliminate the need for optimization of the concentrations of primers, Mg2+, and Taq DNA polymerase, even for difficult assays (e.g., assays in which the copy number of the target gene is much smaller than that for the reference gene).|[A] NH4+ ions prevent nonspecific primers from annealing to the template. [B] Synthetic factor MP, an innovative PCR additive, increases the local concentration of primers and probes at the template. Together with K+ and other cations, synthetic factor MP stabilizes specifically bound primers and probes, allowing efficient primer extension by HotStarTaq Plus DNA Polymerase.|Viral RNA was diluted in serial fivefold dilutions and amplified in duplex with an internal control using the QuantiTect Virus Kit (see figure "Reliable detection of viral RNA over a wide linear range") or kits from Suppliers AII and I. The QuantiTect Virus Kit provided much higher sensitivity than the other reagents tested, enabling reliable analysis of unknown samples. ND: Not detected after 50 PCR cycles.|Serial tenfold dilutions of RNA standards (in vitro transcribed RNA) were prepared using either QuantiTect Nucleic Acid Dilution Buffer or RNase-free water, as indicated. These dilutions were used as template in one-step RT-PCR either directly (0 test) or after storage for 2 or 4 weeks at -20°C. Using QuantiTect Nucleic Acid Dilution Buffer resulted in lower CT values and improved stability of the standards.|
Performance

应用QuantiTect Virus Kits进行扩增可在广泛的稀释范围内获得陡峭的Sigma曲线,即使在模板量少、CT值高的情况下也是如此(参见"Unambiguous determination of CT values over a wide dynamic range")。由此可精确确定real-time PCR的CT值,定量分析病毒核酸。

多重分析可检测含有内参的多种病毒RNA和/或DNA靶分子,适用线性范围广泛,且不影响检测灵敏度(参见"Reliable detection of viral RNA over a wide linear range"和"Improved detection of low amounts of viral RNA")。

试剂盒中提供的QuantiTect Nucleic Acid Dilution Buffer可在稀释和反应体系构建过程中稳定RNA和DNA标准品,避免在试管和枪头等管壁丢失核酸。该缓冲液确保可靠稀释用于病毒核酸定量分析的标准核酸,可获得从低到高CT值的广泛线性范围。缓冲液可避免降解,延长标准品的储存时间(参见"Reliable dilution and storage of RNA standards")。

Principle

QuantiTect Virus Kit可通过单一或多重分析对病毒核酸高度灵敏的检测(参见"QIAGEN multiplex kits")。优化的预混液保证PCR产物在多重反应中的扩增效率和灵敏度与在单一反应中相同。

在同一反应液中扩增内参和靶基因,可减少手工误差,提高基因定量分析的可靠性。QuantiTect Virus Buffer含有浓度平衡的K+、NH4+和独特合成的MP因子,可促使引物和探针与核酸模板稳定、高效的退火,提高PCR效率(参见"Unique PCR buffer")。此外,Sensiscript Reverse Transcriptase确保对病毒RNA高灵敏度的逆转录,而HotStarTaq Plus DNA Polymerase具有严格的热启动,可防止非特异性产物的形成。

QuantiTect Virus Kit的组份
试剂盒组份 特点 优势
5x QuantiTect Virus Master Mix 浓缩的预混液 浓度高,经过优化,可灵敏的检测病毒核酸 起始样本量更大,灵敏度更高
HotStarTaq Plus DNA Polymerase 在95ºC下5分钟即可活化 室温下构建qPCR反应体系
QuantiTect Virus Buffer NH4+和K+的平衡组合 特异性引物退火确保PCR结果可靠
合成的Factor MP 在单管内对4个基因进行可靠的多重分析
其他的试剂盒组份 QuantiTect Virus RT Mix Contains a unique formulation of Sensiscript Reverse Transcriptase 优化用于高度灵敏检测病毒RNA
QuantiTect Nucleic Acid Dilution Buffer Proprietary buffer formulation for dilution and storage of nucleic acid standards. 在稀释和反应体系构建时稳定RNA和DNA标准,避免核酸在塑料表面(如指管或枪头)的损失
Procedure

QuantiTect Virus Kit使用序列特异性探针可对病毒核酸(RNA和/或DNA)及内参进行高度灵敏的real-time PCR分析。反应可选择是否进行逆转录步骤,可灵活设计实验检测RNA、DNA或者RNA及DNA。选用使用手册中的实验方案能快速获得可靠结果。

可选预混液中含有或不含ROX染料的试剂盒(见下表)。

选择合适的QuantiTect Virus Kit
ROX染料试剂盒 兼容的PCR仪
预混液中含有ROX染料 QuantiTect Virus Kit 除Applied Biosystems 7500以外的Applied Biosystems的各种PCR仪
预混液中不含ROX染料 QuantiTect Virus +ROX Vial Kit Applied Biosystems 7500和
Bio-Rad、Cepheid、 Eppendorf、QIAGEN, Roche、Agilent等PCR仪

我们推荐使用QIAGEN OneStep RT-PCR Kit进行快速、高灵敏度的终点式一步法RT-PCR,包括病毒检测。

Applications

QuantiTect Virus Kit可进行高度灵敏的单一或多重real-time PCR,或应用序列特异性探针的一步法RT-PCR,检测内参和病毒DNA和/或RNA。该试剂盒可用于各种real-time PCR仪,包括Applied Biosystems、Bio-Rad、Cepheid、Eppendorf、QIAGEN、Roche(除capillary cyclers以外)和Agilent的PCR仪。

Feature
Specifications
Applications Virus detection
Reaction type Reverse Transcription and PCR
Real-time or endpoint Real-Time
Sample/target type RNA and/or DNA targets
Single or multiplex Single or multiplex
SYBR Green I or sequence-specific probes Sequence-specific probes
Thermal cycler Most real-time cyclers (except capillary cyclers e.g. LightCycler® 1.x and 2.0)
With or without ROX Available with ROX in master mix and ROX as a separate vial

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Brochures & Guides
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Kit Handbooks
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For highly sensitive real-time singleplex or multiplex PCR or one-step RT-PCR using sequence-specific probes for detection of viral DNA and RNA and internal controls
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