Cat Herpes Virus Detection Kit

Cat Herpes Virus Detection KitPackaging specifications 25T/box&50T/box

【 Principle of Inspection 】

Cat Herpes Virus Detection KitThis kit is designed with specific primers and molecular beacon probes for this gene. In the presence of this DNA in the test sample, PCR reaction can proceed and release fluorescent signals. Real time monitoring and output of corresponding fluorescence signals during PCR process using instruments to achieve qualitative analysis of detection results.

Main components

【 Storage conditions and expiration date 】 Store in the dark below -20 ℃ to avoid repeated freezing and thawing, with an expiration date of 12 months.

【 Applicable Instruments 】 Multi channel real-time fluorescence quantitative PCR instruments including ABI series, Bio Rad series, Agilent Stratagene MX series, Roche LightCycler R480, Cepheid SmartCycler, Rotor Gene series, Hangzhou Borui series, etc.

If using the original specimen for testing. The specimens (patient cerebrospinal fluid, venous blood, joint fluid; diseased animal venous blood, breast milk; bacterial colonies, bacterial moss, etc.) should be freshly collected and suspended in sterilized water or sterile physiological saline.

If PCR detection is required after bacterial enrichment, selective enrichment solution should be used to enrich the original specimen.

If the specimen cannot be detected immediately after collection, it can be stored overnight at 2-8 ℃; If long-term storage is required, the specimen should be frozen at -20 ° C to -80 ° C.

1. Reagent Preparation (Reagent Preparation Area)

(1) Take out the reagent kit from the refrigerator, extract the necessary reagents for the experiment from the kit, thoroughly melt and mix, and centrifuge instantly to remove any liquid adhering to the tube wall.

(2) Calculate the required number of reactions (n) for the current experiment, and determine the amount of various reagents needed for the experiment based on the reaction system shown in the table below.

N=number of negative controls (1T)+number of positive controls (1T)+margin of error (1T)+number of samples

(3) Add the above reagents into a sterile centrifuge tube, mix thoroughly, and centrifuge immediately. Divide the reagent into PCR reaction tubes at a rate of 20 μ L per tube.

(4) Cover the PCR reaction tube tightly and transfer it to the sample processing area. Put the remaining reagents back into the refrigerator below -20 ℃ for freezing storage.

2. Sample preparation (sample processing area)

Use DNA/RNA extraction kit to extract nucleic acid, and follow the instructions of the kit for specific operations.

3. Sample addition

Add 5 μ l of processed DNA samples and a final volume of 25 μ l/tube to the prepared PCR reaction sample tubes. Cover the tubes with PCR reaction caps and mix well. Immediately centrifuge at low speed and transfer to the PCR instrument. For the negative control and positive control tubes, add 5 μ L each of the negative control or positive control samples provided in the kit, with a final volume of 25 μ l/tube. Cover the PCR reaction cap and mix well. Immediately centrifuge at low speed and transfer to the PCR instrument.

4. PCR (PCR amplification region)

(1) Take the prepared PCR reaction tubes from the sample processing area, place them in the corresponding position of the real-time fluorescence quantitative PCR instrument sample slot, and record the placement order.

(2) Set the instrument's nucleic acid amplification parameters according to the table below for PCR amplification.

Note: When setting up the ABI series fluorescence PCR instrument, ROX calibration is not selected, and the quenching group is set to None.

Reference value (reference range)

1. Validity assessment of reagent kit:

(1) Positive control: There is a typical S-shaped amplification curve or Ct value ≤ 35.

(2) Negative control: Ct value>38 or no Ct value, linear or slightly diagonal, no exponential growth period.

2. Determination of specimen results:

(1) Positive: The Ct value of the specimen test result is ≤ 35 or there is a significant exponential growth period.

(2) Suspicious: The Ct value of the specimen test result is within the range of 35-38. At this point, the specimen should be subjected to repeated testing. If the Ct value of the repeated experiment is still within the range of 35-38 and there is a significant exponential growth period, it is judged as positive. Otherwise, it is judged as negative.

(3) Negative: The Ct value of the specimen test result is greater than 38 or there is no Ct value.

Explanation of Inspection Results

Limitations of Inspection Methods

When the concentration of the tested nucleic acid in the test sample is lower than the detection of this kit, false negative results may occur.

Improper handling of the collected, transported, stored, and processed samples can easily lead to DNA degradation and false negative results.

3. Cross contamination of samples during collection, transportation, storage, and processing can easily lead to false positive results.

The detection limit of the product is 103 copies/mL, and the CV value of the product is ≤ 3%.

【 Precautions 】

1. Laboratories using this reagent kit should strictly follow the management regulations for gene amplification testing laboratories issued by relevant national departments;

To avoid RNA degradation, the sample processing should be operated under 0-4 ℃ conditions, and the experiment should be immediately tested on the machine after completion; The instruments and consumables used in the sample processing process should be treated with ribonucleases;

3. All items in each area are for exclusive use and must not be used interchangeably to avoid contamination; After the inspection is completed, the workbench should be cleaned immediately;

4. When absorbing the reaction solution, try to avoid generating bubbles as much as possible; Before using the PCR instrument, it is important to check if all reaction tubes are tightly sealed to prevent inaccurate results caused by liquid evaporation;

5. All reagents in the reagent kit should be thoroughly melted and mixed at room temperature before use, and should be centrifuged immediately;

6. The negative and positive controls provided in the reagent kit should be transferred to the specimen preparation area and stored separately before the first use;

To prevent fluorescence interference, direct contact with the PCR reaction tube with bare hands should be avoided, and any labeling on the PCR reaction tube should be avoided;

8. The suction head used during the testing process should be directly poured into a waste tank containing 10% hypochlorous acid. The PCR reaction tube that has been tested should not be opened and should be sterilized together with other waste items before being discarded; Workbenches and various experimental supplies should be regularly disinfected with 10% hypochlorous acid, 75% alcohol, or ultraviolet lamps;

The parameters related to instrument amplification should be set according to the relevant requirements in this manual; Different batch numbers of reagents cannot be mixed; And use it within its validity period. The product is only used for scientific research.