Amplifluor ID Kits

• virus내 RNA로부터 RT-PCR후 Real-time PCR을 하기위해 형광 forward primer, reverse primer를 세트로 제공합니다.
• Real-time fluorescence detection을 통해, 증폭된 물질의 양이 실시간으로 측정됩니다.
• quantitative or qualitative evaluation에 사용됩니다.
• hairpin primer를 이용하여 sensitivity를 높여줍니다.

Primer 구조

원리

Figure 1. The Amplifluor® Primer
The Amplifluor® system consists of the fluorescent Amplifluor® hairpin primer and an unconjugated reverse primer. In combinations, these two primers produce a fluorescently labeled amplicon, which can be measured by real-time PCR (right). The carefully designed Amplifluor® molecule consists of four parts - a fluorophore, a hairpin tructure, a DABSYL quencher, and a targetspecific sequence tail (top). The Amplifluor® primer has little fluorescence in the native closed state (energy transfer), but upon incorporation into an amplicon during PCR, he hairpin unfolds, separating the fluorochrome and quencher creating a substantial signal which can be easily detected (right).

실험방법

종류
• Amplifluor® ID Avian Influenza A (H5N1) Kit
  Influenza serotype인 H5N1 viruse의 hemagglutinin (H5)또는 neuraminidase (N1)를 encoding하는 genome내 conserved nucleic acid를, 두종류의 hairpin구조의 primer를 이용하여 Real time PCR을 함으로서 influenza를 쉽게 측정할 수 있는 제품입니다.
  • 제품구성
    • 25x Amplifluor® ID Influenza A, H5 Forward Primer (Part No. 2005861)
    • 25x Amplifluor® ID Influenza A H5 Reverse Primer (Part No. 2005863)
    • 25x Amplifluor® ID Influenza A N1 Forward Primer (Part No. 2005865)
    • 25x Amplifluor® ID Influenza A N1 Reverse Primer (Part No. 2005867)
    • N1 Positive Control (Part No. AMP3500-1)
    • H5 Positive Control (Part No. AMP3500-2)
• Amplifluor® ID Herpes Virus Type 1 (HSV-1) & Type 2 (HSV-2) Kit
  glycoprotein B (gB), extracellular tropism receptor of HSV-1 (HSV-1 primers)와 glycoprotein D (gD), extracellular tropism receptor of HSV-2 (HSV-2 primers)를 이용하여 Real-time PCR하여 Herpes Virus Type 1과 Type 2를 검출할 수 있습니다.
  • 제품구성
    • 25x Amplifluor® ID HSV-1 Forward Primer (Part No. 2005849)
    • 25x Amplifluor® ID HSV-1 Reverse Primer (Part No. 2005851)
    • 25x Amplifluor® ID HSV-2 Forward Primer (Part No. 2006942)
    • 25x Amplifluor® ID HSV-2 Reverse Primer (Part No. 2006943)
    • HSV-1 Positive Control (Part No. AMP3800-1)
    • HSV-2 Positive Control (Part No. AMP3800-2)
• Amplifluor® ID Pan-Enterovirus Detection Kit
  일반적인 enteroviral strain의 5’ UTR을 특이적으로 인식하는 primer를 이용하여 Real time PCR함으로서 enterovirus를 검출할 수 있습니다.
  • 제품구성
    • 25x Amplifluor® ID Pan-Enterovirus Forward Primer (Part No. 2003585)
    • 25x Amplifluor® ID Pan-Enterovirus Reverse Primer (Part No. 2003586)
    • Pan-enterovirus Positive Control (Part No. AMP3100-1)
• Amplifluor® ID West Nile Virus Detection Kit
  West Nile Virus의 NS-5 nucleic acid region을 인식하도록 설계된 primer를 이용하여 Real time PCR함으로서 west Nile Virus RNA의 정량 또는 정성 분석 할 수 있습니다.
  • 제품구성
    • 25x Amplifluor® ID West Nile Forward Primer (Part No. 2003793)
    • 25x Amplifluor® ID West Nile Reverse Primer (Part No. 2003792)
    • West Nile Virus Positive Control (Part No. AMP3200-1)

사용예

A.

B.

C.

Evaluation of the Amplifluor® ID Human Herpesvirus Type 1 (HSV-1) Detection Kit. Ten-fold serial dilutions of HSV-1 control was amplified using the Amplifluor® ID HSV-1 primer set and Bio-RadTM iTaq® Supermix 2X Master Mix analyzed in triplicate. Samples were run on the ABI 7500® Real-Time PCR instrument using the following parameters: 95ºC for 3 minutes; followed by 50 cycles of 95ºC for 15 seconds and 60ºC for 35 seconds. The mean CT value of diluted control plasmid was plotted on the Y axis against the Log 10 (copy number of oncentration) on the X axis. The PCR efficiency (Ex) of HSV-1 control plasmid was determined using the CT slope method with 5 data points from a 5 log dilution range (Figure 1C). The calculated efficiency for this assay is 97% with an R2 value of 0.9941. The detection of the no template control (NTC) was not observed up to the late 46 cycles (Figure 1. A, B & C). The data below is for reference use only and the data should not be used to interpret actual assay results.

• Herpesviruses Detection Kit