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Enterovirus 71 Nucleic Acid Detection Kit(Enzymatic Probe Isothermal Amplification)

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Enterovirus 71 Nucleic Acid Detection Kit(Enzymatic Probe Isothermal Amplification)

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Introduction

[Product Name] Enterovirus 71 Nucleic Acid Detection Kit(Enzymatic Probe Isothermal Amplification)

[Packaging Size] 50 tests/kit

[Intended Use]

This kit is intended for in vitro qualitative detection of Enterovirus 71 nucleic acid in human throat swab samples.

Hand-foot-and-mouth disease(HFMD) is an acute infectious disease caused by enterovirus infection. At present, a total of 108 enterovirus serotypes have been found, which are divided into four groups: A, B, C and D. The disease mostly occurs in children under 5 years old, and can cause herpes on the hands, feet, mouth and other parts, and also can cause complications such as myocarditis, pulmonary edema, aseptic meningoencephalitis of a small number of children . There are more than 20 types of enteroviruses that cause HFMD, among which enterovirus 71 (EV71) is main pathogens causing HFMD in children.

[Test Principle]

The kit was to detect nucleic acid of EV71 using Enzymatic Probe Isothermal Amplification (EPIA). Primers and Probe with RNA bases (rProbe) were developed tailored to the highly conservative sequences in genomes of EV71 and added Bst enzyme and RNaseH. Two ends of RNA bases in rProbe were labeled with fluorophores as reporter and quencher. During amplification, the target sequence was amplified in isothermal condition using DNA polymerase and strand replacement of Bst enzyme, RNaseH cleaves RNA bases on target gene-probe hybrid strands, enabling to separate the reporter dye from the quencher dye and to generate the fluorescent signal. The remaining segment of rProbe RNA can continue the amplification process to generate fluorescent signals further. The fluorescent signals can be monitored by detection system, thereby realizing the detection of the target nucleic acid.

Recombinant pseudo-virus of Pine chloroplast gene was developed as internal control in the kit to monitor the quality in laboratory processing including validity of reagent and laboratory operation to avoid false negative.

[Storage Condition and Shelf-life]

The kit can be stored below -18℃ and protected from light. The shelf life is 9 months. Repeated freezing and thawing cannot exceed 4 cycles. It can be transported stably for 5 days below -18°C and protected from light.

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

[Applicable Instruments]

Applied Biosystems 7500 Real-Time PCR Systems, SLAN ® -96P Real-Time PCR Systems, LightCycler® 480 Real-Time PCR system, Easy Amp Real-time Fluorescence Isothermal Detection System(HWTS1600)

[Requirements for Samples]

  1. Sample Collection

Freshly collected throat swabs.

Wipe the posterior pharyngeal wall and bilateral tonsils at the back of the uvula (palatine or uvula) with moderate force with a polyester fiber head swab, avoid touching the tongue, take out the posterior sampling tube, and break the plastic handle at the contact part of the hand, soak the swab into the sampling buffer, and tighten the cap. Throat swab samples can be preserved with a preservative solution such as normal saline or Hank's solution. If the Macro & Micro-Test Sample Release Reagent (HWTS-3005-8) from Jiangsu Macro & Micro-Test Med-Tech Co., Ltd. is used, the collected swab can be directly put into the sample release reagent.

  1. Sample Storage

The collected samples should be tested as soon as possible, and the samples that cannot be tested within 24 hours should be stored in a cryopreservation tube immediately after collection, marked with the sample name (or serial number) and type, and stored below -70°C. Samples should be stored separately in a special library or a special counter.

  1. Sample Transportation

It is not more than 5 days for sealed transportation in foam box with dry ice, and not more than 3 days for transportation with ice bags.

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