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Detection Kit for Infectious Bursal Disease Virus RNA(PCR-Fluorescence Probing)
【Product Name】Real-time PCR Detection Kit for Infectious Bursal Disease Virus
【Package】50 kits/box
【Indication】The Real-time PCR Detection Kit for Infectious Bursal Disease Virus is applicable to detect the Infectious Bursal Disease Virus RNA in avian throat swab, cloaca swab, tissue, chicken embryo allantoic fluid and cell culture. The test results are for research purpose only and not for clinical diagnosis.
【Main components and content】
Name | Specification | Quantity |
IBDV Reaction Solution | 1000µl/Tube | 1 |
IBDV 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 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 2µl nucleic acid of negative control, positive control and nucleic acid samples into the above PRC reaction tubes respectively, centrifuge at 8000rpm for several seconds, and put them into the fluorescent quantitative PCR amplifier.
3. qR-T-PCR amplification
3.1 Selection of Fluorescence channel
Reporter Dye1: select the FAM channel to detect MS-H vaccine strain, Quencher Dye: NONE
Reporter Dye2: select the VIC or HEX channel to detect the MS wild virus strain, Quencher Dye2: NONE, passive reference: NONE. Refer to the instrument manual for other instruments.
3.2 The reaction conditions are set as:
Parameters of amplification | |||
System | Total volume is 25µl | ||
Reaction Conditions of PCR | Phase | Conditions | Cycles |
Pre-denaturation | 95℃ for 5 minutes | 1 | |
PCR | 95℃ for 10 seconds |
40 | |
60℃ for 30 seconds | |||
【Interpretation of the results】
1. The following requirements shall be matched at the same time in one experiment, otherwise the experiment is invalid and needs to be repeated.
Negative control had no amplification curve
The amplification curve of positive control FAM channel had a significant logarithmic growth phase, and the Ct value of the FAM channel amplification curve was ≤ 32.
2. If the FAM channel of the test sample has no amplification curve, the sample can be determined as IBDV negative. If the FAM channel has a logarithmic growth phase amplification curve and the Ct value ≤ 36, which can be determined as IBDV positive. 36 < Ct value < 40 are suspicious samples, which need to be retested for confirmation.
【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
Detection Kit for Infectious Bursal Disease Virus RNA(PCR-Fluorescence Probing)
【Product Name】Real-time PCR Detection Kit for Infectious Bursal Disease Virus
【Package】50 kits/box
【Indication】The Real-time PCR Detection Kit for Infectious Bursal Disease Virus is applicable to detect the Infectious Bursal Disease Virus RNA in avian throat swab, cloaca swab, tissue, chicken embryo allantoic fluid and cell culture. The test results are for research purpose only and not for clinical diagnosis.
【Main components and content】
Name | Specification | Quantity |
IBDV Reaction Solution | 1000µl/Tube | 1 |
IBDV 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 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 2µl nucleic acid of negative control, positive control and nucleic acid samples into the above PRC reaction tubes respectively, centrifuge at 8000rpm for several seconds, and put them into the fluorescent quantitative PCR amplifier.
3. qR-T-PCR amplification
3.1 Selection of Fluorescence channel
Reporter Dye1: select the FAM channel to detect MS-H vaccine strain, Quencher Dye: NONE
Reporter Dye2: select the VIC or HEX channel to detect the MS wild virus strain, Quencher Dye2: NONE, passive reference: NONE. Refer to the instrument manual for other instruments.
3.2 The reaction conditions are set as:
Parameters of amplification | |||
System | Total volume is 25µl | ||
Reaction Conditions of PCR | Phase | Conditions | Cycles |
Pre-denaturation | 95℃ for 5 minutes | 1 | |
PCR | 95℃ for 10 seconds |
40 | |
60℃ for 30 seconds | |||
【Interpretation of the results】
1. The following requirements shall be matched at the same time in one experiment, otherwise the experiment is invalid and needs to be repeated.
Negative control had no amplification curve
The amplification curve of positive control FAM channel had a significant logarithmic growth phase, and the Ct value of the FAM channel amplification curve was ≤ 32.
2. If the FAM channel of the test sample has no amplification curve, the sample can be determined as IBDV negative. If the FAM channel has a logarithmic growth phase amplification curve and the Ct value ≤ 36, which can be determined as IBDV positive. 36 < Ct value < 40 are suspicious samples, which need to be retested for confirmation.
【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
