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Freeze-dried Adenovirus Nucleic Acid Detection Kit (Enzymatic Probe Isothermal Amplification)

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Freeze-dried Adenovirus Nucleic Acid Detection Kit (Enzymatic Probe Isothermal Amplification)

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Introduction

[Product Name] Freeze-dried Adenovirus Nucleic Acid Detection Kit (Enzymatic Probe Isothermal

Amplification)
[Packaging Size] 48 tests/kit
[Intended Use]
This kit is used for in vitro qualitative detection of Adenovirus nucleic acid in nasopharyngeal swab and throat swab samples.
Human adenovirus (HAdV) is a common pathogen that can cause acute respiratory infectious diseases in humans, mainly through droplet transmission and contact transmission. Adenoviruses are widely distributed in nature and have many types. There are 7 groups (A-G), and there are 55 serotypes having pathogenic effects on humans, among which causing the respiratory tract infections in humans are mainly group B (types 3, 7, 11, 14, 16, 21, 50, 55), Group C (Types 1, 2, 5, 6, 57) and Group E (Type 4) [1-8]. Different types have different clinical symptoms, but mainly respiratory tract infections. Respiratory diseases caused by respiratory tract infections of human body account for 5%~15% of global respiratory diseases, and 5%~7% of global childhood respiratory diseases [9], which can also cause acute gastroenteritis, epidemic keratoconjunctivitis, etc.. Adenovirus is endemic in a wide range of areas and can be infected all year round, especially in crowded areas, which are prone to local outbreaks, mainly in schools and military camps. Due to the lack of corresponding protective antibodies, children are susceptible to infection, leading to severe pneumonia, which is the focus of respiratory adenovirus infection.
The test results of this kit are not the sole indicator for the diagnosis of adenovirus infection in patients. It must be combined with the clinical symptoms of the patient and other laboratory test data to correctly determine the infection of the pathogen, and formulate a reasonable treatment plan to make the treatment safe and effective.

[Test Principles]

This kit adopts Enzymatic Probe Isothermal Amplification (EPIA), and designs specific primers and RNA base-containing probes (rProbe) for the highly conserved region of ADV, and adds Bst enzyme and RNaseH enzyme at the same time, in which the left and right ends of the RNA base of rProbe are labeled with fluorescent groups and quencher, respectively. Use the DNA polymerase activity and strand displacement activity of Bst enzyme to amplify the target to be tested at a constant temperature, the RNaseH enzyme can cleave the RNA bases on the target-probe hybrid

chain, so that the fluorescent group and quencher of rProbe are separated thereby fluorescing. In addition, the left fragment of the residual rProbe RNA base can be used as a primer to further extend to form a product, which further accumulates the product. The fluorescent signal is continuously accumulated with the formation of the product, thereby realizing the detection of the target nucleic acid.

In the system, the recombinant pine chloroplast gene plasmid is used as an exogenous internal control, as the quality control of reagent and operation itself, to avoid false negative.

[Storage Conditions and Shelf-life]

1. Unopened kit: It should be stored at no higher than 30°C and protected from light, and the shelf-life is 12 months.
2. It should be tested immediately after the freeze-dried reaction buffer, positive control and internal control are reconstituted. The remaining reconstituted positive control and internal control should be stored below -15°C away from light, and the shelf-time is 1 week and the repeated freezing and thawing should be avoided (no more than 2 cycles).

3. Transportation conditions: It can be transported stably for 1 month below 37°C.

4. See the packaging label for the production date, production batch number and expiry date.

[Applicable Instruments]

Applied Biosystems 7500 Real-Time PCR Systems, SLAN-96P Real-Time PCR Systems (Shanghai Hongshi Medical Technology Co., Ltd.), LightCycler®480 Real-Time PCR system, Real-time Fluorescence Constant Temperature Detection System Easy Amp HWTS1600.

[Requirements for Samples]

1. Sample Collection
1.1 Nasopharyngeal swab
Use a swab with the tip of polyester fiber to enter from the anterior nostril, and slowly go deep backward along the bottom of the inferior meatus. When the top of the swab reaches the posterior wall of the nasopharyngeal cavity (feeling against the wall), place the swab still for a moment (about 3 seconds), then gently rotate it for a round, slowly remove the swab, and put it into the tube. Break the plastic handle by hand, immerse the swab in the sampling solution, and tighten the tube cap. It has been proved that nasal swab samples can be preserved in preservation solutions such as saline or Hank’s solution.
1.2 Throat swab
Use polyester fiber head swab to wipe the posterior uvula, posterior pharyngeal wall and bilateral tonsils repeatedly with moderate force, and avoid touching the tongue. Take out the rear sampling tube, break the plastic handle on the contact part of the hand, soak the swab in the sampling solution, and tighten the tube cap. It has been proved that throat swab samples can be preserved in preservation solutions such as saline or Hank’s solution.
2. Storage
The samples to be tested should be stored at 2-8°C for no more than 7 days, stored below -18°C for no more than 4 months, and stored below -70°C for long term. The samples should be avoided from repeated freezing and thawing.

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