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Detection Kit for Avian Influenza Virus H5 Subtype RNA(PCR-Fluorescence Probing)
（Product Name）Detection Kit for Avian Influenza Virus H5 Subtype RNA(PCR-Fluorescence Probing)
（Package）50 kits/box
（Indication）The Detection Kit for Avian Influenza Virus H5 Subtype RNA(PCR-Fluorescence Probing) is applicable to detect the Avian Influenza virus H5 subtype 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 |
H5-AVI Reaction Solution | 1150µl/Tube | 1 |
H5-AVI Positive Control | 100µl/Tube | 1 |
Negative Control | 100µl/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 23µl to of reaction solution to each tube.
2.2 Add 2µl 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 fluorescent quantitative PCR amplifier.
2.3 The probe detection mode is set as: Reporter Dye1: FAM, and Quencher Dye:NONE.
2.4 The reaction conditions are set as follows:
55℃ for 15 minutes, one cycle; 95℃ for 30s, one cycle; 95℃ for 10s → 60℃ for 30s (collect the fluorescence), 40 cycles. Save the file and run it.
（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 or the Ct value > 40, the sample can be determined as Avian Influenza H5 subtype negative. If the FAM channel has a logarithmic growth phase amplification curve and the Ct value ≤ 40, which can be determined as Avian Influenza H5 subtype positive.
（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 Avian Influenza Virus H5 Subtype RNA(PCR-Fluorescence Probing)
（Product Name）Detection Kit for Avian Influenza Virus H5 Subtype RNA(PCR-Fluorescence Probing)
（Package）50 kits/box
（Indication）The Detection Kit for Avian Influenza Virus H5 Subtype RNA(PCR-Fluorescence Probing) is applicable to detect the Avian Influenza virus H5 subtype 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 |
H5-AVI Reaction Solution | 1150µl/Tube | 1 |
H5-AVI Positive Control | 100µl/Tube | 1 |
Negative Control | 100µl/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 23µl to of reaction solution to each tube.
2.2 Add 2µl 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 fluorescent quantitative PCR amplifier.
2.3 The probe detection mode is set as: Reporter Dye1: FAM, and Quencher Dye:NONE.
2.4 The reaction conditions are set as follows:
55℃ for 15 minutes, one cycle; 95℃ for 30s, one cycle; 95℃ for 10s → 60℃ for 30s (collect the fluorescence), 40 cycles. Save the file and run it.
（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 or the Ct value > 40, the sample can be determined as Avian Influenza H5 subtype negative. If the FAM channel has a logarithmic growth phase amplification curve and the Ct value ≤ 40, which can be determined as Avian Influenza H5 subtype positive.
（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
