HPAI Quadrivalent PCR

Detection Kit for Avian Influenza Virus H5/H7/H9/General Subtype RNA(PCR-Fluorescence Probing)

【Product Name】

Real-Time PCR Detection Kit for Avian Influenza Virus H5/H7/H9/General Subtype

【Package】

50 kits/box

【Main components and content】

【Storage and Shelf Life】

Stored at -20±5℃, Repeated freezing and thawing ≤ 3 times, the shelf life is 12 months.

【Model】

ABI 7500, ABI QuantStudio 5, CFX Connect, CFX Opus 96, LightCycler480, Gentiter 96E/96R, LineGene 9600 Plus and other fluorescent quantitative PCR amplifier.

【Test Method】

1. Nucleic Acid Extraction

Commercialized RNA extraction kit can be carried out for nucleic acid extraction, please follow the instruction of the kit.

2. PCR amplification

2.1 Calculate the number of test samples, take n+2 PCR reaction tubes, and add 25µl to of reaction solution to each tube.

2.2 Add 5µl nucleic acid of negative control, positive control and samples into the above PCR reaction tubes respectively, centrifuge at 8000rpm for several seconds, and put them into the fluorescent quantitative PCR amplifier..

2.3 The reaction conditions are set as follows:

【Interpretation of the results】

1. Determination of test kit effectiveness :

(1) Weak positive control: Ct value of FAM, HEX,Cy5 and ROX channels ≤ 32, amplification curve with obvious exponential phase.

(2) Blank control: FAM, HEX, Cy5 and ROX channels have no amplification curve, or the amplification curve is straight or slight oblique, no significant exponential phase, and Ct value ≥ 38 or no Ct value.

2. Determination of the results:

*Note:

(1) If there is a logarithmic growth phase amplification curve and Ct value ≤ 36, it is judged as +. If there is no amplification curve or Ct value > 36, it is judge as -. The samples are suspicious when 36 < Ct value <40, which has to be retested.

(2) If the results of any H5/H7/H9 subtype is + with ROX channel -, it has to be retested.

【Precautions】

1. The laboratory management shall be in strict accordance with the management specification of PCR gene amplification laboratory. The laboratory personnel must be trained professionally. The experiment process shall be conducted strictly in different areas (Reagent preparation area, specimen preparation area, amplification and product analysis area). All consumables shall be disposable after sterilization. Special apparatus, equipment and supplies at each stage of the experiment operation shall not be cross-used.

2. Please prepare the biological safety cabinet for reagent and specimen preparation stage. The Lab coat, disposable gloves and pipettor shall be carried out during the experiment.

3. Repeated freezing&thawing of reagents shall be avoided as far as possible. Before use, the reagents shall be completely thawed and centrifuged at 8000rpm for several seconds.

4. Please put the pipette used in the specimen preparation area into the container containing disinfectant and discard with the waste after sterilization.

5. After the experiment, the worktable and pipettor were treated with 10% hypochlorite or 75% alcohol or ultraviolet lamp.

【Manufacture】

Name: Shandong Xinda Gene Technology Co., Ltd

A subsidiary of the Shandong Sinder Technology Co., Ltd

Address: Building B2, Bandaohuigu Industrial Park, Shungeng Road, Zhucheng City, Shandong Province

Post Code: 262233

Phone: +86 - 0532 5882 0810