PCR Detection Kit for Enterocytozoon Hepatopenaei Disease (EHP) of Shrimp (Fluorescent Probe Assays)
Product Name: Real-time PCR Detection Kit for Enterocytozoon Hepatopenaei Disease.
Package: 50 kits/box
Indication: This kit is applicable to the nucleic acid detection of Enterocytozoon Hepatopenaei Disease (EHP) DNA in shrimp muscle tissue The test results are for research purpose only and not for clinical diagnosis.
Main components and content:
| Name |
Specification |
Quantity |
| EHP Reaction Solution |
1000µl/Tube |
1 |
| EHP Positive Control |
250µl/Tube |
1 |
| Negative Control |
250µl/Tube |
1 |
Storage and Shelf Life:
Stored at -20±5ºC,Repeated freezing and thawing ≤ 3 times, the shelf life is 12 months.
Test Method:
- Nucleic Acid Extraction
Commercialized DNA extraction kit can be carried out for nucleic acid extraction, please follow the instruction of the kit.
- 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 fluorescent quantitative PCR.
2.3 The reaction conditions are set as:
| Parameters of amplification |
| System |
Total volume is 25µl |
| Signal collection |
WSSV fluorescence signal |
FAM channel collects fluorescence signal |
Reaction Conditions of PCR |
Phase |
Conditions |
Cycles |
| UNG Process |
37ºC for 2 minutes |
1 |
| Pre-denaturation |
95ºC for 30 seconds |
1 |
PCR |
95ºC for 10 seconds |
40 |
60ºC for 30 seconds
Set to collect the fluorescent signal at the end of this phase |
Result Judgement:
- The validity judgement:
- Positive Control: The Ct value of FAM channel ≤ 32, and the amplification curve has a significant exponential growth phase.
- Negative Control: FAM channel has not amplification curve, or the amplification curve is straight line or slight oblique line, with no significant exponential growth phase, and the Ct value ≥ 38 or no Ct value.
- The test result judgement:
- If there is a significant exponential growth in FAM channel and Ct value ≤35, it is be judged as EHP positive.
- If there is a significant exponential growth in FAM channel and Ct value between 35-38, it is be judged as EHP suspicious. The sample should be tested repeatedly. If the Ct value of the repeated test results is still between 35-38 with an significant exponential growth period, it is judged as positive, otherwise it is negative.
- If the Ct value of FAM test results > 38 or no Ct value, it is judged as EHP negative
Precautions:
- 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.
- 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.
- 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.
- Please put the pipette used in the specimen preparation area into the container containing disinfectant and discard with the waste after sterilization.
- 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
Shandong Sinder Technology Co., Ltd ('Sinder') founded in April 1999, has a wide range of animal health products. Sinder focus on Animal Health Products, Animal Vaccine, Antibodies, Antibiotics, Feed Additives, Chinese Herbal Product, Pet food, Elisa, and PCR test kit.
Sinder always persists in the core concept of 'The Relentless Pursuit of Increasing Customer Value. ' Based on the animal health to achieve a service provider for ecological construction of agriculture and animal husbandry. Support the efficient and sustainable development of agriculture and animal husbandry. Make food safer and rural areas better!
Sinder is a high-tech enterprise and one of the top 10 veterinary drug manufacturers in China and focusing on the animal health business. At the moment, the company has 35 GMP production lines, 1 CNAS laboratory, 3 national scientific research platforms, and 8 provincial scientific research platforms owned by Sinder.
Sinder has 4 R&D centers located in Qingdao, Zhucheng, and Beijing, China, and 1 laboratory in Silicon Valley in the United States. At present, the company has 1,535 employees including more than 200 masters or above and 18 doctors. Also, the company established extensive technical cooperation with several domestic and foreign scientific research institutes, animal health companies, and domestic veterinary medicine enterprises. Sinder has undertaken 57 national, provincial, and ministerial projects, gained 7 provincial and ministerial science and technology progress awards, presided over or participated in 23 national standards, obtained 23 new veterinary drug certificates(one 1st class new drug), and 60 authorized patents.
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