Biokey Super Sensitive Multiplex Real Time PCR Kit Nocardia seriolae Nucleic Acid Detection Kit

Intended use:

The kit is used for the epidemiological analysis of Nocardia seriolae in fish viscera tissue samples

Principle:

In this kit, the specific fragment of N.seriolae is used to design the primer probe probe, which can specifically bind to a SECTION of DNA template in the middle of the primer amplification region. During the PCR extension process, the exonuclide activity of Taq enzyme cuts the 5 '-terminal fluorophore from the probe and makes it free in the reaction system. Therefore, the quenched 3 '-end group was separated and the fluorescence was emitted. The fluorescence PCR instrument was used to detect Nocardia seriolae in a completely closed reaction system

Main components:

1. Sample type

Gills, spleen, kidney and other pathological tissues.

2. Sample collection and transportation

Samples shall be collected and transported in accordance with SC/T 7103. Samples shall be transported to the laboratory as soon as possible under refrigerated conditions to avoid repeated freeze-thaw.

3. Sample preservation

It can be stored for 24 hours at 2℃~8℃, and can be stored for a long time at -20℃.

PCR amplification and fluorescence detection (PCR detection area):

Put each reaction tube into the PCR instrument and perform PCR amplification according to the following procedures:

FAM was selected as the detection channel.

Quality control:

1. Negative control: No obvious amplification curve of FAM detection channel;
2. Positive control: FAM channel showed obvious amplification curve, Ct value ≤30;
3. The above conditions are met simultaneously in the same experiment; otherwise, the experiment is invalid and re-testing is required.

The results of interpretation

Note: The result is judged to be grey area, which needs to be rechecked. If the recheck result is Ct value < 45 is considered as positive, and no Ct value or Ct value ≥45 is considered as negative.

Product performance index:

1. Accuracy

The positive reference materials of testing enterprises were all positive.

2. Minimum detection limit: the detection limit of this kit is 10copies/μL.

3. Precision: Coefficient of variation (CV,%) of intra-batch precision Ct value is ≤5%.

4. Linear, linear correlation coefficient | r | > 0.980.

Product picture:

Operation details please check from IFU!