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Detection Kit for Avian Influenza Virus H5/H7/Universal Subtype RNA(PCR-Fluorescence Probing)
【Product Name】Detection Kit for Avian Influenza Virus H5/H7/Universal Subtype RNA(PCR-Fluorescence Probing)
【Package】50 kits/box
【Indication】The Detection Kit for Avian Influenza Virus H5/H7/Universal RNA(PCR-Fluorescence Probing) is applicable to detect the Avian Influenza virus H5/H7/Universal 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/H7/Universal Trivalent Reaction Solution | 1250µl/Tube | 1 |
H5/H7/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.
【Model】
ABI 7500, ABI QuantStudio 5, CFX Connect, CFX Opus 96, LightCycler480, Gentiter 96E/96R, LineGene 9600 Plus and other fluorescent quantitative PCR amplifier.
【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 25µl 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 fluorescent quantitative PCR amplifier..
2.3 The reaction conditions are set as follows:
Relevant parameters of the amplifier | |||
System | Total volume: 30µl | ||
Signal collection | Avian Influenza H5/H7/Universal | H5 subtype - HEX channel collects fluorescence signal | |
H7 subtype - FAM channel collects fluorescence signal | |||
Universal - ROX 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 | |
56℃: 30 seconds (Set to collect fluorescent signal at the end of this stage) |
【Interpretation of the results】
1. Determination of test kit effectiveness :
(1) Weak positive control: Ct value of FAM, HEX and ROX channels ≤ 32, amplification curve with obvious exponential phase.
(2) Blank control: FAM, HEX and ROX channels have no amplification curve, or the amplification curve is straight or slight oblique, no significant exponential phase, and Ct value ≥ 38 or no Ct value.
2. Determination of the results:
Result Judgement | FAM channel | HEX channel | ROX channel |
Avian influenza virus H5 subtype nucleic acid positive | - | + | + |
Avian influenza virus H7 subtype nucleic acid positive | + | - | + |
Low pathogenic avian influenza virus nucleic acid positive | - | - | + |
Avian influenza virus H5/H7 nucleic subtype acid positive | + | + | + |
Avian influenza virus nucleic acid negative | - | - | - |
*Note:
(1) If there is a logarithmic growth phase amplification curve and Ct value ≤ 36, it is judged as +. If there is no amplification curve or Ct value > 36, it is judge as -. The samples are suspicious when 36 < Ct value <40, which has to be retested.
(2) If the results of any H5/H7 subtype is + with ROX channel -, it has to be retested.
【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/H7/Universal Subtype RNA(PCR-Fluorescence Probing)
【Product Name】Detection Kit for Avian Influenza Virus H5/H7/Universal Subtype RNA(PCR-Fluorescence Probing)
【Package】50 kits/box
【Indication】The Detection Kit for Avian Influenza Virus H5/H7/Universal RNA(PCR-Fluorescence Probing) is applicable to detect the Avian Influenza virus H5/H7/Universal 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/H7/Universal Trivalent Reaction Solution | 1250µl/Tube | 1 |
H5/H7/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.
【Model】
ABI 7500, ABI QuantStudio 5, CFX Connect, CFX Opus 96, LightCycler480, Gentiter 96E/96R, LineGene 9600 Plus and other fluorescent quantitative PCR amplifier.
【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 25µl 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 fluorescent quantitative PCR amplifier..
2.3 The reaction conditions are set as follows:
Relevant parameters of the amplifier | |||
System | Total volume: 30µl | ||
Signal collection | Avian Influenza H5/H7/Universal | H5 subtype - HEX channel collects fluorescence signal | |
H7 subtype - FAM channel collects fluorescence signal | |||
Universal - ROX 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 | |
56℃: 30 seconds (Set to collect fluorescent signal at the end of this stage) |
【Interpretation of the results】
1. Determination of test kit effectiveness :
(1) Weak positive control: Ct value of FAM, HEX and ROX channels ≤ 32, amplification curve with obvious exponential phase.
(2) Blank control: FAM, HEX and ROX channels have no amplification curve, or the amplification curve is straight or slight oblique, no significant exponential phase, and Ct value ≥ 38 or no Ct value.
2. Determination of the results:
Result Judgement | FAM channel | HEX channel | ROX channel |
Avian influenza virus H5 subtype nucleic acid positive | - | + | + |
Avian influenza virus H7 subtype nucleic acid positive | + | - | + |
Low pathogenic avian influenza virus nucleic acid positive | - | - | + |
Avian influenza virus H5/H7 nucleic subtype acid positive | + | + | + |
Avian influenza virus nucleic acid negative | - | - | - |
*Note:
(1) If there is a logarithmic growth phase amplification curve and Ct value ≤ 36, it is judged as +. If there is no amplification curve or Ct value > 36, it is judge as -. The samples are suspicious when 36 < Ct value <40, which has to be retested.
(2) If the results of any H5/H7 subtype is + with ROX channel -, it has to be retested.
【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