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Real-Time PCR Detection Kit for African Swine Fever Virus (MGF/CD2V/VP72)
【Product Name】
Real-time PCR Detection Kit for African Swine Fever Virus (MGF/CD2V/VP72)
【Package】
50 kits/box
【Main components and content】
Name | Specification | Quantity |
ASFV (MGF/CD2V/VP72) Reaction Solution | 1000µl/Tube | 1 |
ASFV (MGF/CD2V/VP72) Positive Control | 250µl/Tube | 1 |
Negative Control | 250µl/Tube | 1 |
【Storage and Shelf Life】
Stored at -20±5℃
,Repeated freezing and thawing ≤ 3 times, the shelf life is 12 months.
【Test Method】
1. Nucleic Acid Extraction
Commercialized DNA 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 20µl to of reaction solution to each tube.
2.2 Add 5µl l 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:
Relevant parameters of the amplifier | |||
System | Total volume: 30µl | ||
Signal collection | ASFV (MGF/CD2V/VP72) Fluorescent Signal | VP72-FAM channel collects fluorescence signal | |
CD2V-VIC/HEX channel collects fluorescence signal | |||
MGF-CY5 channel collects fluorescence signal | |||
PCR reaction conditions | Stage | Condition | Cycle number |
UNG processing | 37℃: 2 minutes | 1 | |
Predegeneration | 95℃: 30 seconds | 1 | |
PCR | 95℃: 10 seconds | 40 | |
56℃: 30 seconds (Set to collect fluorescent signal at the end of this stage) |
【Interpretation of the results】
1. Determination of test kit effectiveness :
(1) Positive control: Ct value of FAM, HEX/VIC and CY5 channels ≤ 32, amplification curve with obvious exponential phase.
(2) Negative control: FAM, HEX/VIC and CY5 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:
Result Judgement | FAM channel | HEX /VIC channel | CY5 channel |
VP72 ASFV nucleic acid positive | + | - | - |
CD2V ASFV nucleic acid positive | - | + | - |
MGF ASFV nucleic acid positive | - | - | + |
VP72 and CD2V ASFV nucleic acid positive | + | + | - |
VP72 and MGF ASFV nucleic acid positive | + | - | + |
CD2V and MGF ASFV nucleic acid positive | - | + | + |
MGF, CD2V and VP72 ASFV nucleic acid positive | + | + | + |
MGF, CD2V and VP72 ASFV nucleic acid negative | - | - | - |
*Note:
(1) If there is a logarithmic growth phase amplification curve and Ct value ≤ 35, it is judged as +. If there is no amplification curve or Ct value > 38, it is judge as -. The samples are suspicious when 35 < Ct value <38, which has to be retested. If the Ct value of retested result is still between 35-38 with obvious logarithmic growth phase, it is judged as positive, otherwise negative.
【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
Real-Time PCR Detection Kit for African Swine Fever Virus (MGF/CD2V/VP72)
【Product Name】
Real-time PCR Detection Kit for African Swine Fever Virus (MGF/CD2V/VP72)
【Package】
50 kits/box
【Main components and content】
Name | Specification | Quantity |
ASFV (MGF/CD2V/VP72) Reaction Solution | 1000µl/Tube | 1 |
ASFV (MGF/CD2V/VP72) Positive Control | 250µl/Tube | 1 |
Negative Control | 250µl/Tube | 1 |
【Storage and Shelf Life】
Stored at -20±5℃
,Repeated freezing and thawing ≤ 3 times, the shelf life is 12 months.
【Test Method】
1. Nucleic Acid Extraction
Commercialized DNA 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 20µl to of reaction solution to each tube.
2.2 Add 5µl l 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:
Relevant parameters of the amplifier | |||
System | Total volume: 30µl | ||
Signal collection | ASFV (MGF/CD2V/VP72) Fluorescent Signal | VP72-FAM channel collects fluorescence signal | |
CD2V-VIC/HEX channel collects fluorescence signal | |||
MGF-CY5 channel collects fluorescence signal | |||
PCR reaction conditions | Stage | Condition | Cycle number |
UNG processing | 37℃: 2 minutes | 1 | |
Predegeneration | 95℃: 30 seconds | 1 | |
PCR | 95℃: 10 seconds | 40 | |
56℃: 30 seconds (Set to collect fluorescent signal at the end of this stage) |
【Interpretation of the results】
1. Determination of test kit effectiveness :
(1) Positive control: Ct value of FAM, HEX/VIC and CY5 channels ≤ 32, amplification curve with obvious exponential phase.
(2) Negative control: FAM, HEX/VIC and CY5 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:
Result Judgement | FAM channel | HEX /VIC channel | CY5 channel |
VP72 ASFV nucleic acid positive | + | - | - |
CD2V ASFV nucleic acid positive | - | + | - |
MGF ASFV nucleic acid positive | - | - | + |
VP72 and CD2V ASFV nucleic acid positive | + | + | - |
VP72 and MGF ASFV nucleic acid positive | + | - | + |
CD2V and MGF ASFV nucleic acid positive | - | + | + |
MGF, CD2V and VP72 ASFV nucleic acid positive | + | + | + |
MGF, CD2V and VP72 ASFV nucleic acid negative | - | - | - |
*Note:
(1) If there is a logarithmic growth phase amplification curve and Ct value ≤ 35, it is judged as +. If there is no amplification curve or Ct value > 38, it is judge as -. The samples are suspicious when 35 < Ct value <38, which has to be retested. If the Ct value of retested result is still between 35-38 with obvious logarithmic growth phase, it is judged as positive, otherwise negative.
【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