Yellow Head Virus (YHV) Nucleic Acid Detection Kit (RT-PCR-Fluorescent Probe Method)

Intended use:

This kit is suitable for the specific detection of pathogenic yellow head virus (YHV) in tissue samples such as gills, antennae glands, hematopoietic tissues and lymphatic organs of penaeus prawn.

Principle:

This kit uses RNA as a template and primers as a starting point to synthesize cDNA strands complementary to RNA template under the action of high-efficiency reverse transcriptase. Choose cat goblet virus gene conservative design specific probe, the probe can with primer amplification area in the middle of a specific binding DNA template, in the process of PCR extension, the circumscribed Taq polymerase enzymes from the 5 'end of fluorescent groups on the probe will be cut down, make it free in the reaction system, thereby quenching groups out of the 3' end, glow, The nucleic acid of Prawn yellow head virus (YHV) was detected by fluorescence PCR in a completely closed reaction system.

Main components:

1. Sample type

Tissue samples from prawn

2. Sample collection

Samples were collected according to SNT 1151.4-2003.

Take fresh prawn tissue as sample.

2.1 For larvae or young shrimp, whole shrimp or shrimp heads are taken as samples. To remove the head, use sterilizing scissors and tweezers to remove the eyes, breastplate, all thoracic appendages, and abdomen behind the head.

2.2 For adult shrimp, gill filaments, lymphatic organs or hemolymph were taken as samples.

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 is selected for detection channels.

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.

Operation details please check from IFU!

Bio key Health is a high-tech enterprise with integrating R&D, production and sales&marketing in Guangzhou. Since 2003. Bio Key was founded by the "Thousand Talents Program" entrepreneurial team and the overseas senior automation equipment development engineer team.

The company's core team has been in the field of IVD research and entrepreneurship for more than 10 years, and has mastered a wealth of molecular diagnostic POCT industrial application technology, SmartTRF immune detection technology, RT qPCR nucleic acid detection technology and independent research and development capabilities of instruments.

It has a number of independent core technologies in the fields of automation engineering, micro-precision manufacturing, immunofluorescence, nucleic acid detection and gene sequencing. Since its establishment, the company has applied for more than 60 patents and currently has 17 authorized patents. The company has two R&D centers for diagnostic reagents and instruments and equipment, totaling more than 500 square meters; it has established more than 2,000 square meters of GMP workshops and equipment production plants; it has SmartTRF new immune detection technology, nucleic acid detection technology (POCT) and three scientific services.