For highly sensitive detection of viral RNA and/or DNA
- High sensitivity in single and multiplex assays
- Detection of viral RNA and/or DNA in the same reaction
- Clear detection of weak positive signals
- Fast universal 2-step protocols
- 5x master mix for higher sensitivity with more sample input
The QuantiTect Virus Kit is specially designed for highly sensitive detection of up to 4 viral nucleic acid targets by multiplex, real-time one-step RT-PCR using sequence-specific probes. Multiple viral RNA and/or DNA targets plus internal controls can be detected in a single tube without loss of sensitivity. The concentrated 5x master mix enables the addition of higher sample input and contains HotStarTaq Plus DNA Polymerase. The QuantiTect Virus RT Mix contains a unique formulation of Sensiscript Reverse Transcriptase that is optimized for highly sensitive detection of viral RNA. Two kit formats are available: the QuantiTect Virus Kit for cyclers that require ROX dye for fluorescence normalization and the QuantiTect Virus +ROX Vial Kit for other cyclers. For convenience, the master mix can be stored at 2–8°C.
nn-NO
Configure
Added to your shopping cart
|
Product
|
Product no.
|
Cat. no.
|
List price:
|
|
|
This product has been discontinued.
Instead, we recommend
|
|
Show details
|
|
|
varies
|
select
Can't order online?
To place an order via phone, email or for requesting a quote, please provide the product’s name, number and catalog number.
|
|
|
| Product |
Cat. no. |
List price: |
|
|
Product
|
Cat. no.
|
List price:
|
|
|
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
|
|
|
|
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
|
|
|
|
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
|
|
|
|
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
|
|
|
|
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
|
|
|
QuantiTect Virus Kits are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.
幅広いダイナミックレンジで明確なCT 値を測定可能|幅広い範囲で直線性のあるウイルスRNA検出|QIAGEN multiplex kits|ユニークなPCRバッファー|従来のマルチプレックスよりも低濃度でウイルスRNAを検出|RNAスタンダードの正確な希釈および保存|
ウシウイルス性下痢ウイルス(BVDV)のRNA連続希釈液(図参照)をQuantiTect Virus Kitを用いて増幅し、解析した。シャープなS状曲線により低コピーのテンプレート量でも正確なC T 値を決定できた(10 copies/μl)。 [A] BVDV1 RNAの検出。 [B] BVDV2 RNAの検出。NTC:No Template Control。|ウイルスRNAの連続5倍希釈溶液を調製し、インターナルコントロールと共にQuantiTect Virus Kitを用いてduplexで増幅した。 [A] 標的ウイルスの増幅プロット。 [B] インターナルコントロールの増幅プロット。|QuantiTect Multiplex PCR Kit は簡単な操作でマルチプレックスリアルタイム定量PCRを実現する。汎用されている方法に比べて、本キットはプライマー、Mg 2+、 Taq DNA polymeraseの濃度についての至適化が不要で、リファレンス遺伝子に比べてターゲット遺伝子のコピー数が少ないアッセイでも同様。| [A] NH 4+ イオンは非特異的なプライマーのテンプレートへのアニーリングを防ぐ。 [B]画期的なPCR添加物であるFactor MP は、テンプレート周辺でのプライマー濃度を高める。K + や他の陽イオンと共に、Factor MPは特異的に結合したプライマーを安定化し、HotStarTaq Plus DNA Polymeraseによる効果的なプライマーエクステンションを可能にする。|ウイルスRNAを5倍連続希釈し、インターナルコントロールとともにQuantiTect Virus Kitを用いてduplexで増幅 (図 " 幅広い範囲で直線性のあるウイルスRNA検出")、または Suppliers A II および I社の製品を用いて増幅した。The QuantiTect Virus Kitは他の試薬よりもはるかに感度が高く、未知のサンプルでも信頼できる解析が可能である。 ND:PCRで50サイクル後でも検出できなかった。|QuantiTect Nucleic Acid Dilution Buffer またはRNase フリー水を用いて、表記のようにRNA スタンダード(in vitro 転写RNA)の10 倍連続希釈溶液を調製した。これらの希釈液を調製直後(0 test)、あるいは-20℃で2 および4 週間保存後に1 ステップRT-PCR のテンプレートとして用いた。QuantiTect Nucleic Acid Dilution Buffer を用いた場合のほうが、低いC T 値が得られ、スタンダードの安定性も高い結果が得られた。|
パフォーマンス
Amplification using QuantiTect Virus Kits provides steep sigmoidal curves over a range of dilutions, even for low template amounts with high CT values (see figure "Unambiguous determination of CT values over a wide dynamic range"). This enables accurate CT value determination for quantification of viral nucleic acids in real-time PCR.
Multiplex assays enable the detection of multiple viral RNA and/or DNA targets plus internal controls over a wide linear range without loss of sensitivity (see figures "Reliable detection of viral RNA over a wide linear range" and "Improved detection of low amounts of viral RNA").
QuantiTect Nucleic Acid Dilution Buffer, supplied with the kits, stabilizes RNA and DNA standards during dilution and reaction setup and prevents loss of nucleic acids on plastic surfaces, such as tubes or pipet tips. The buffer enables reliable dilution of standards used to quantify viral nucleic acids, giving a wide linear range, from low to high CT values and ensures longer storage of standards without degradation (see figure "Reliable dilution and storage of RNA standards").
原理
QuantiTect Virus Kits provide highly sensitive detection of viral nucleic acids in single or multiplex assays on the first attempt (see flowchart "QIAGEN multiplex kits"). The optimized master mix ensures that PCR products in a multiplex reaction are amplified with the same efficiency and sensitivity as PCR products in a corresponding single-amplification reaction.
Amplifying control and target genes in the same reaction, instead of separate reactions, increases the reliability of gene quantification by minimizing handling errors. The QuantiTect Virus Buffer contains a balanced combination of K+ and NH4+ ions as well as unique synthetic Factor MP stabilizes, which together promote stable and efficient annealing of primers and probes to the nucleic acid template, enabling high PCR efficiency (see figure "Unique PCR buffer"). In addition, the unique formulation of Sensiscript Reverse Transcriptase ensures highly sensitive reverse transcription of viral RNA, while HotStarTaq Plus DNA Polymerase provides a stringent hot start, preventing the formation of nonspecific products.
| 5x QuantiTect Virus Master Mix |
Concentrated master mix |
Highly concentrated and optimized for sensitive virus detection |
Larger volumes of template can be added to the assay for increased sensitivity |
| HotStarTaq Plus DNA Polymerase |
5 min activation at 95ºC |
Set up of qPCR reactions at room temperature |
| QuantiTect Virus Buffer |
Balanced combination of NH4+ and K+ ions |
Specific primer annealing ensures reliable PCR results |
| Synthetic Factor MP |
Reliable multiplexing analysis of up to 4 genes in the same tube |
| Additional kit components |
QuantiTect Virus RT Mix |
Contains a unique formulation of Sensiscript Reverse Transcriptase |
Optimized for highly sensitive detection of viral RNA |
| QuantiTect Nucleic Acid Dilution Buffer |
Proprietary buffer formulation for dilution and storage of nucleic acid standards. |
Stabilizes RNA and DNA standards during dilution and reaction setup and prevents loss of nucleic acids on plastic surfaces, such as tubes or pipet tips |
操作手順
QuantiTect Virus Kits provide highly sensitive real-time PCR analysis of viral nucleic acids (RNA and/or DNA) and internal controls using sequence-specific probes. Reactions can be carried out with or without a reverse-transcription step, enabling flexible design of multiplex assays to detect RNA targets, DNA targets, or both RNA and DNA targets. Follow the protocol in the handbook for fast and reliable results.
Kits are available with or without ROX passive reference dye in the master mix (see table).
| Supplied in master mix |
QuantiTect Virus Kit |
All cyclers from Applied Biosystems except Applied Biosystems 7500 |
| Supplied in separate tube |
QuantiTect Virus +ROX Vial Kit |
Applied Biosystems 7500 and cyclers from Bio-Rad, Cepheid, Eppendorf, QIAGEN, Roche, Agilent, and other suppliers |
For fast and highly sensitive end-point one-step RT-PCR applications, including virus detection, we recommend using the QIAGEN OneStep RT-PCR Kit.
アプリケーション
QuantiTect Virus Kits provide highly sensitive real-time single or multiplex PCR or one-step RT-PCR using sequence-specific probes for detection of viral DNA and/or RNA plus internal controls. The kits can be used on a wide range of real-time thermal cyclers, including instruments from Applied Biosystems, Bio-Rad, Cepheid, Eppendorf, QIAGEN, Roche (except for capillary cyclers), and Agilent.
|
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
|
|
|
|
|
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
|
Show details
|
Images
Unambiguous determination of CT values over a wide dynamic range.
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.
Reliable detection of viral RNA over a wide linear range.
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.
QIAGEN multiplex kits.
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).
Unique PCR buffer.
[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.
Improved detection of low amounts of viral RNA.
Viral RNA was diluted in serial fivefold dilutions and amplified in duplex with an internal control using the QuantiTect Virus Kit (see previous 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.
Reliable dilution and storage of RNA standards.
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.
|