 |
Real-time PCR Detection Kit for Newcastle Disease Virus Virulent and Universal Strains
For Veterinary Use Only
（Product Name）Real-time PCR Detection Kit for Newcastle Disease Virus Virulent and Universal Strains.
（Package）50 Tests
（Indication）The Detection Kit for Newcastle Disease Virus Virulent and Universal Strains is applicable to detect the NDV Virulent/Universal Strains RNA in bird throat swabs, cloaca swabs, tissue and culture cell samples. The test results are for research purposes only and not for clinical diagnosis.
（Main components and content）
Name | Specification | Quantity |
NDV Virulent/Universal Reaction Solution | 1000µl/Tube | 1 |
NDV Virulent/Universal 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
A commercialized RNA/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 put them into the Thermal Cycler.
2.3 The reaction conditions are set as follows:
Relevant parameters of the amplifier | |||
System | Total volume: 25µl | ||
Signal collection | NDV Virulent/Universal Fluorescence signals | Virulent - FAM channel collects fluorescence signal | |
Universal – HEX channel collects fluorescence signal | |||
PCR reaction conditions | Stage | Condition | Cycle number |
UNG process | 50℃: 5 minutes | 1 | |
Predegeneration | 95℃: 30 seconds | 1 | |
PCR | 95℃: 10 seconds |
40 | |
58℃: 45 seconds (Set to collect fluorescent signal at the end of this stage) | |||
*Note: Please do not choose ROX correction for ABI series fluorescence quantitative Thermal Cycler, choose “None” for Quenchers.
（Interpretation of the results）
1. Determination of test kit effectiveness:
1.1 Positive Control: The Ct values of the FAM and HEX channel ≤ 32, and the amplification curve has a significant exponential growth phase.
1.2 Negative Control: FAM and HEX channel have no amplification curve, or the amplification curve is a straight line or slightly oblique line.
2. Determination of the results:
Result Judgement | FAM channel | HEX channel |
NDV Virulent Strain nucleic acid positive | + | + |
NDV Non-Virulent Strain (Vaccine) nucleic acid positive | - | + |
NDV nucleic acid negative | - | - |
*Note:
If there is an Exponential growth amplification curve and Ct value ≤ 36, it is determined as “+”.
If there is no amplification curve, it is determined as “-”.
If 36 < Ct Value < 40, it is a doubtful sample, and the analysis should be repeated for confirmation.
（Precautions）
1. The laboratory management shall strictly follow the management specification of the 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 pipette 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 it with the waste after sterilization.
5. After the experiment, the worktable and pipette shall be 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 Newcastle Disease Virus Virulent and Universal Strains
For Veterinary Use Only
（Product Name）Real-time PCR Detection Kit for Newcastle Disease Virus Virulent and Universal Strains.
（Package）50 Tests
（Indication）The Detection Kit for Newcastle Disease Virus Virulent and Universal Strains is applicable to detect the NDV Virulent/Universal Strains RNA in bird throat swabs, cloaca swabs, tissue and culture cell samples. The test results are for research purposes only and not for clinical diagnosis.
（Main components and content）
Name | Specification | Quantity |
NDV Virulent/Universal Reaction Solution | 1000µl/Tube | 1 |
NDV Virulent/Universal 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
A commercialized RNA/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 put them into the Thermal Cycler.
2.3 The reaction conditions are set as follows:
Relevant parameters of the amplifier | |||
System | Total volume: 25µl | ||
Signal collection | NDV Virulent/Universal Fluorescence signals | Virulent - FAM channel collects fluorescence signal | |
Universal – HEX channel collects fluorescence signal | |||
PCR reaction conditions | Stage | Condition | Cycle number |
UNG process | 50℃: 5 minutes | 1 | |
Predegeneration | 95℃: 30 seconds | 1 | |
PCR | 95℃: 10 seconds |
40 | |
58℃: 45 seconds (Set to collect fluorescent signal at the end of this stage) | |||
*Note: Please do not choose ROX correction for ABI series fluorescence quantitative Thermal Cycler, choose “None” for Quenchers.
（Interpretation of the results）
1. Determination of test kit effectiveness:
1.1 Positive Control: The Ct values of the FAM and HEX channel ≤ 32, and the amplification curve has a significant exponential growth phase.
1.2 Negative Control: FAM and HEX channel have no amplification curve, or the amplification curve is a straight line or slightly oblique line.
2. Determination of the results:
Result Judgement | FAM channel | HEX channel |
NDV Virulent Strain nucleic acid positive | + | + |
NDV Non-Virulent Strain (Vaccine) nucleic acid positive | - | + |
NDV nucleic acid negative | - | - |
*Note:
If there is an Exponential growth amplification curve and Ct value ≤ 36, it is determined as “+”.
If there is no amplification curve, it is determined as “-”.
If 36 < Ct Value < 40, it is a doubtful sample, and the analysis should be repeated for confirmation.
（Precautions）
1. The laboratory management shall strictly follow the management specification of the 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 pipette 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 it with the waste after sterilization.
5. After the experiment, the worktable and pipette shall be 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
