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Real-Time PCR Detection Kit for African Swine Fever Virus (VP72/I177L)
【Product Name】Real-time PCR Defection Kit for African Swine Fever Virus (VP72/I177L)
【Package】50 kits/box
【Indication】The Detection Kit for African Swine Fever Virus is applicable to detect the African Swine Fever Virus VP72/I177L Subtype DNA in Pig saliva, blood, tissue and environmental samples. The test results are for research purpose only and not for clinical diagnosis
【Main components and content】
Name | Specification | Quantity |
1000µl/Tube | 1 | |
ASFV (VP72/I177L) 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.
【Application】ABI QuantStudio 5, CFX Opus 96, LightCycler 480, Gentier 96E/96R, LineGene 9600 Plus and other real-time PCR instruments.
【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 nucleic acid of negative control, positive control and samples into the above PRC reaction tubes respectively, centrifuge at 8000rpm for several seconds, and place them into the Real-Time PCR...
2.3 The reaction conditions are set as follows:
Relevant parameters of the amplifier | |||
System | Total volume: 25µl | ||
Signal collection | ASFV (VP72/I177L) Fluorescent Signal | I177L-HEXchannel collects fluorescence signal | |
VP72-FAM 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 | |
60℃: 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 and HEX channels ≤ 32, amplification curve with obvious exponential phase.
(2) Negative control: FAM and HEX channels have no amplification curve, or the amplification curve is straight or slight oblique.
2. Determination of the results:
Result Judgement | FAM channel | HEX channel |
- | + | |
VP72 ASFV nucleic acid positive | + | - |
VP72 and I177L ASFV nucleic acid positive | + | + |
VP72 and I177L ASFV nucleic acid negative | - | - |
*Note:
(1) If there is an exponential growing amplification curve and Ct value ≤ 36, it can be determined as “+”. If there is no amplification curve, it can be determined as “-”. When 36 < Ct value <40, samples should be considered as doubtful results, and the analysis should be repeated for conformation.
【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 (VP72/I177L)
【Product Name】Real-time PCR Defection Kit for African Swine Fever Virus (VP72/I177L)
【Package】50 kits/box
【Indication】The Detection Kit for African Swine Fever Virus is applicable to detect the African Swine Fever Virus VP72/I177L Subtype DNA in Pig saliva, blood, tissue and environmental samples. The test results are for research purpose only and not for clinical diagnosis
【Main components and content】
Name | Specification | Quantity |
1000µl/Tube | 1 | |
ASFV (VP72/I177L) 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.
【Application】ABI QuantStudio 5, CFX Opus 96, LightCycler 480, Gentier 96E/96R, LineGene 9600 Plus and other real-time PCR instruments.
【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 nucleic acid of negative control, positive control and samples into the above PRC reaction tubes respectively, centrifuge at 8000rpm for several seconds, and place them into the Real-Time PCR...
2.3 The reaction conditions are set as follows:
Relevant parameters of the amplifier | |||
System | Total volume: 25µl | ||
Signal collection | ASFV (VP72/I177L) Fluorescent Signal | I177L-HEXchannel collects fluorescence signal | |
VP72-FAM 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 | |
60℃: 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 and HEX channels ≤ 32, amplification curve with obvious exponential phase.
(2) Negative control: FAM and HEX channels have no amplification curve, or the amplification curve is straight or slight oblique.
2. Determination of the results:
Result Judgement | FAM channel | HEX channel |
- | + | |
VP72 ASFV nucleic acid positive | + | - |
VP72 and I177L ASFV nucleic acid positive | + | + |
VP72 and I177L ASFV nucleic acid negative | - | - |
*Note:
(1) If there is an exponential growing amplification curve and Ct value ≤ 36, it can be determined as “+”. If there is no amplification curve, it can be determined as “-”. When 36 < Ct value <40, samples should be considered as doubtful results, and the analysis should be repeated for conformation.
【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
