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Freeze-dried Coxsackie Virus Type A16 Nucleic Acid Detection Kit (Enzymatic Probe Isothermal Amplification)

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Freeze-dried Coxsackie Virus Type A16 Nucleic Acid Detection Kit (Enzymatic Probe Isothermal Amplification)

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Introduction

[Product Name] Freeze-dried Coxsackie Virus Type A16 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 Coxsackie virus type A16 nucleic acid in human throat swabs.

Hand-foot-mouth disease (HFMD) is mainly an acute infectious disease caused by enterovirus infection. The disease mostly occurs in children under 5 years old and could cause herpes on the parts such as hands, feet, mouth, etc., and complications such as myocarditis, pulmonary edema, sterile meningoencephalitis, etc. could be caused in a few child patients. There are more than 20 types of enteroviruses that cause HFMD, of which Coxsackie virus type A16 (Cox A16) is one of the main pathogens causing HFMD in child patients.

[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 Cox A16, 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 reconstituted reaction buffer 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
Freshly collected throat swabs.
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. If use the Macro & Micro-Test Sample Release Reagent (HWTS-3005-8), the collected swabs could be directly put into the sample release reagent.
2. Storage
The collected samples should be sent for test as soon as possible. Specimens that cannot be tested within 24 hours should be placed in cryopreservation tubes immediately after collection, marked with the sample name (or serial number) and type, and stored below -70°C. Specimens should be stored separately in a special warehouse or counter.
3. Transport
It is not more than 5 days for transportation in sealed foam boxes added with dry ices, and not more than 3 days for low temperature transportation in ice packs.

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