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Real-time PCR Detection Kit for Porcine Epidemic Diarrhea, Transmissible Gastroenteritis and Porcine Deltacoronavirus Disease Virus
[Product Name] Real-time PCR Detection Kit for Porcine Epidemic Diarrhea, Transmissible Gastroenteritis and Porcine Deltacoronavirus Disease Virus
[Package] 50 Tests
[Indication] The Real-time PCR Detection Kit for Porcine Epidemic Diarrhea, Transmissible Gastroenteritis and Porcine Deltacoronavirus Disease Virus is applicable for detection, typing, auxiliary diagnosis and epidemiological investigation of the PED/TGEV/PDCoV in porcine serum, plasma, intestinal tissue, intestinal contents, mesenteric lymph nodes, excreta and other samples. The test results are for research purpose only and not for clinical diagnosis.
[Main components and content]
Name | Specification | Quantity |
PED/TGEV/PDCoV Reaction Solution | 1000µl/Tube | 1 |
PED/TGEV/PDCoV 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.
[Model]
For ABI QuantStudio 5、CFX Opus 96、LightCycler480、Gentier 96E/96R、LineGene 9600 Plus and other thermal cyclers.
[Test Method]
1. Nucleic Acid Extraction
A commercialized DNA/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 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: 30µl | ||
Signal collection | Porcine Epidemic Diarrhea (PED) - FAM channel collects fluorescence signal | ||
Transmissible Gastroenteritis (TGEV) – HEX/VIC channel collects fluorescence signal | |||
Porcine Deltacoronavirus Disease Virus (PDCoV) - Cy5 channel collects fluorescence signal | |||
PCR reaction conditions | Stage | Condition | Cycle number |
Reverse Transcription | 55℃: 15 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, 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.
2. Determination of the results:
Result Judgement | FAM channel | HEX/VIC channel | Cy5 channel |
PED acid positive | + | - | - |
TGEV nucleic acid positive | - | + | - |
PDCoV nucleic acid positive | - | - | + |
PED and TGEV nucleic acid positive | + | + | - |
PED and PDCoV nucleic acid positive | + | - | + |
TGEV and PDCoV nucleic acid positive | - | + | + |
PED, TGEV and PDCoV nucleic acid positive | + | + | + |
PED, TGEV and PDCoV 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 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 the 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 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 Porcine Epidemic Diarrhea, Transmissible Gastroenteritis and Porcine Deltacoronavirus Disease Virus
[Product Name] Real-time PCR Detection Kit for Porcine Epidemic Diarrhea, Transmissible Gastroenteritis and Porcine Deltacoronavirus Disease Virus
[Package] 50 Tests
[Indication] The Real-time PCR Detection Kit for Porcine Epidemic Diarrhea, Transmissible Gastroenteritis and Porcine Deltacoronavirus Disease Virus is applicable for detection, typing, auxiliary diagnosis and epidemiological investigation of the PED/TGEV/PDCoV in porcine serum, plasma, intestinal tissue, intestinal contents, mesenteric lymph nodes, excreta and other samples. The test results are for research purpose only and not for clinical diagnosis.
[Main components and content]
Name | Specification | Quantity |
PED/TGEV/PDCoV Reaction Solution | 1000µl/Tube | 1 |
PED/TGEV/PDCoV 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.
[Model]
For ABI QuantStudio 5、CFX Opus 96、LightCycler480、Gentier 96E/96R、LineGene 9600 Plus and other thermal cyclers.
[Test Method]
1. Nucleic Acid Extraction
A commercialized DNA/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 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: 30µl | ||
Signal collection | Porcine Epidemic Diarrhea (PED) - FAM channel collects fluorescence signal | ||
Transmissible Gastroenteritis (TGEV) – HEX/VIC channel collects fluorescence signal | |||
Porcine Deltacoronavirus Disease Virus (PDCoV) - Cy5 channel collects fluorescence signal | |||
PCR reaction conditions | Stage | Condition | Cycle number |
Reverse Transcription | 55℃: 15 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, 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.
2. Determination of the results:
Result Judgement | FAM channel | HEX/VIC channel | Cy5 channel |
PED acid positive | + | - | - |
TGEV nucleic acid positive | - | + | - |
PDCoV nucleic acid positive | - | - | + |
PED and TGEV nucleic acid positive | + | + | - |
PED and PDCoV nucleic acid positive | + | - | + |
TGEV and PDCoV nucleic acid positive | - | + | + |
PED, TGEV and PDCoV nucleic acid positive | + | + | + |
PED, TGEV and PDCoV 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 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 the 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 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