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Freeze-dried Influenza B Virus Nucleic Acid Detection Kit(Enzymatic Probe Isothermal Amplification)

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Freeze-dried Influenza B Virus Nucleic Acid Detection Kit(Enzymatic Probe Isothermal Amplification)

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Introduction

[Product Name]

Freeze-dried Influenza B Virus Nucleic Acid Detection Kit(Enzymatic Probe Isothermal Amplification)
[Packaging Size]
48 tests/kit

[Intended Use]

This kit in intended for in vitro qualitative detection of Influenza B virus nucleic acid in nasopharyngeal and oropharyngeal swab samples.
Influenza virus, a representative species of Orthomyxoviridae, is a pathogen that seriously threatens human health and can widely infect hosts. Seasonal influenza epidemics infect about 600 million people worldwide and cause 250,000 to 500,000 deaths every year , of which influenza B virus is one of the main causes[1]. Influenza B virus, also known as IVB, is a single-stranded negative-stranded RNA. According to the nucleotide sequence of its antigenic characteristic HA1 region, it can be divided into two major lineages, the representative strains are B/Yamagata/16/88 and B/Victoria /2/87(5)[2]. Influenza B virus generally has strong host specificity. It has been found that IVB can only infect humans and seals, and generally does not cause a worldwide pandemic, but it can cause regional seasonal epidemics [3]. Influenza B virus can be transmitted through various routes such as the digestive tract, respiratory tract, skin damage and conjunctiva. The symptoms are mainly high fever, cough, runny nose, myalgia, etc. Most of them are accompanied by severe pneumonia, severe heart attack. In severe cases, heart, kidney and other organ failure leads to death, and the fatality rate is very high [4]. Therefore, there is an urgent need for a simple, accurate and rapid method for detecting influenza B virus, which can provide guidance for clinical medication and diagnosis.

The test results are not the only basis for the diagnosis of influenza B virus infection, and a comprehensive analysis needs to be carried out in combination with the clinical manifestations of patients and other test results.
[Test Principle]

The kit was to detect nucleic acid of IVB using Enzymatic Probe Isothermal Amplification (EPIA). Primers and Probe with RNA bases (rProbe) were developed tailored to the highly conservative sequences in genomes of IVB and added Bst enzyme and RNaseH. Two ends of RNA bases in rProbe were labeled with fluorophores as reporter and quencher. During amplification, the extracted RNA was reverse transcribed into cDNA, 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 including validity of reagent and laboratory operation to avoid false negative.

[Storage Condition 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.Test should be performed immediately after reconstitution of the freeze-dried reaction buffer, internal control and positive control, the remaining internal control and positive control should be stored below -15°C and protected from light. The validity period is 1 week, and repeated freezing and thawing should be avoided ( limited to 2 cycles).
3.Transportation conditions: It can be transported stably for 1 month when the temperature is not higher than 37°C.
4. 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
1.1 Nasopharyngeal swab
A swab with a polyester fiber tip is used to enter through the anterior nostril and proceed slowly back and forth along the base of the inferior meatus. When the top of the swab reaches the posterior wall of the nasopharyngeal cavity (with a touch of the wall), let the swab stand for a while (about 3 seconds), then gently rotate it for one round, slowly remove the swab, and put it into the tube. Break the plastic handle, immerse the swab in the sampling buffer, and tighten the cap. Nasopharyngeal swab samples can be preserved in normal saline or Hank's solution.
1.2 Oropharyngeal swab
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.

2. Storage
The collected samples should be tested within 4 hours, otherwise, the sample can be stored at 2~8°C for no more than 1 week. If the sample can not be tested with in 24 hours, it should be stored below -18°C for no more than 3 months. The samples can be stored for a long time below -70°C.

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