2019-nCov Automated Extraction and Detection Solution by TIANGEN

Date: Feb 29 2020 Browse: Source: TIANGEN

In December 2019, a series of pneumonia cases of unknown cause has started from Wuhan, Hubei Province, and soon spread to most provinces and cities in China, and many other countries in January 2020. As of 22:00 pm on January 27, 28 confirmed cases and 5794 suspected 2019-nCov cases have been reported in China. The infection source of the virus was inferred as Rhinolophus, and the present mortality rate is 2.9%.

On January 12, 2020, the World Health Organization (WHO) announced that the pneumonia epidemic was caused by a new type of coronavirus (2019-nCov). Coronavirus is a kind of virus that spreads among animals. The virus nucleic acid type is single-stranded RNA. At the same time, WHO also released the nucleic acid sequence information of nCov shared by Chinese scholars, which has provided the molecular detection basis for virus detection and has made the quick detection and identification of pathogens possible.

WHO suggests that people who have acute respiratory diseases such as cough, fever, respiratory tract infection and have been to Wuhan within 14 days or have been exposed to contact with other patients should be comprehensively tested. On 17 January 2020, WHO released the “Laboratory testing for 2019 novel coronavirus (2019-nCoV) in suspected human cases, Interim guidance, 17 January 2020”. The guidance points out that the specimens to be collected from symptomatic patients include respiratory specimens (nasopharynx and oropharyngeal swabs, sputum, bronchoalveolar lavage, etc.) and serum specimens, as follows:

*For transport of samples for viral detection, use VTM (viral transport medium) containing antifungal and antibiotic supplements. For

bacterial or fungal culture, transport dry or in a very small amount of sterile water. Avoid repeated freezing and thawing of specimens.

Aside from specific collection materials indicated in the table also assure other materials and equipment are available: e.g. transport

containers and specimen collection bags and packaging, coolers and cold packs or dry ice, sterile blood-drawing equipment(e.g. needles,

syringes and tubes), labels and permanent markers, PPE, materials for decontamination of surfaces.

For virus detection, serum specimens can be used to detect virus by immunological method, while RT-qPCR is recommended to detect nCov nucleic acid for faster and more accurate detection. As the virus sequence is known, the content of viral nucleic acid in the specimen can be detected quickly by only selecting appropriate reagents and matching primers.

  (Diagnostic detection of Wuhan coronavirus 2019 by real-time RTPCR, Protocol and preliminary evaluation as of Jan 13, 2020)

As a leader in the field of nucleic acid extraction and detection in China, TIANGEN BIOTECH (BEIJING) CO., LTD. has provided hand-foot-mouth virus and influenza A (H1N1) virus detection reagents for more than 10 million people. In 2019, TIANGEN’s virus nucleic acid extraction and detection reagents have been applied to 30 million African classical swine fever samples, making outstanding contributions to the diagnosis and prevention of African classical swine fever in China.

TIANGEN not only provides rapid and accurate virus extraction and detection reagents, but also provides convenient and efficient tissue grinding instruments and automatic nucleic acid extractors, which constitute a complete set of solutions for virus extraction and detection.

● Automated Nucleic Acid Extraction Solution●

TIANGEN Automated Nucleic Acid Extractors are automatic platforms for nucleic acid extraction using magnetic bead method. The application of these platforms not only greatly reduces the workload of inspection and quarantine departments, but also reduces the operational errors of manual extraction, and ensures the consistency and stability of the quality of extracted nucleic acid.

TIANGEN Automated Nucleic Acid Extractor has various throughputs (including 16, 24, 32, 48, 96 channels), and the matching reagents can be used for the nucleic acid extraction from a variety of sample types. TIANGEN also provides customized reagent development and instrument integration services according to different experimental requirements.

1.TGrinder H24 Tissue Homogenizer

● Widely used for grinding and homogenizing various tissues and feces samples

● With grinding homogenization force 2-5 times that of traditional instruments

● Uniform grinding and homogenization, avoiding cross contamination

● Automatic protection device to protect the safety of laboratory personnel

2.TGuide S32 Automated Nucleic Acid Extractor

● Sample throughput: 1-32 samples

● Processing volume: 20-1000 μl

● Sample type: Blood, cells, tissue, feces, virus and other samples

● Processing time: Up to 8 min to obtain viral nucleic acid

● Control mode: Dual control mode of Windows Pad and screen button

● Platform development: Open platform, free to match reagents

3.TGuide S32 Magnetic Viral DNA/RNA Kit (DP604)

● Wide application: High-quality virus DNA/RNA can be purified from serum, plasma, swab sample, tissue treatment solution and various virus preservation solutions.

● Simple and efficient: This product is perfectly matched with TGuide S32 Automated Nucleic Acid Extractor, which can extract virus DNA/RNA with high yield, high purity, stable and reliable quality.

● Downstream applications: The purified nucleic acid is suitable for downstream experiments of virus PCR and real-time PCR.

● Manual Nucleic Acid Extraction Solution●

As a leader in the field of nucleic acid extraction and detection, TIANGEN possesses the most comprehensive series of nucleic acid extraction products in China, which are ideal for coronavirus detection from all kinds of samples: blood, serum/plasma, tissue, swab, virus, etc.

TIANamp Virus DNA/RNA Kit (DP315)

● High efficiency: High quality virus DNA and RNA can be obtained by rapid purification with high repeatability

● High purity: Completely removal of pollutants and inhibitors for downstream application

● High safety: Organic reagent extraction or ethanol precipitation are not needed

● RNA Virus Detection Solution ●

1. TEasy Automated Pipetting System

● High accuracy: The cooling block can keep the temperature of reagent sample below 7 ℃ for more than 60 min. Each APM is calibrated according to international standards, which has higher precision than manual pipetting.

● Easy operation: Small size. Light weight. No tool is needed for block replacement. Built-in PCR/qPCR preparation procedure. Simplified manual operation of PCR solution preparation

● Wide application: Can be used for 96/384-well plate pipetting, PCR, qPCR, gene detection and other high-throughput experiments

2. FastKing One Step RT-qPCR Kit (Probe)(FP314)

The FastKing One Step RT-qPCR Kit (Probe) (FP314) developed by TIANGEN is a one-step reverse transcription fluorescence quantitative detection kit based on probe method, which is specially designed for the detection of trace genes in various samples. The King RTase in the kit is a new molecular modified reverse transcriptase, which has stronger RNA affinity and thermal stability, with improved reverse transcription efficiency and the extension ability of complex secondary structure RNA templates. A new hot start Taq DNA polymerase is also applied to give the PCR reaction higher amplification efficiency and specificity. In addition, the kit simplifies the components to the greatest extent by mixing Taq and MLV enzyme into enzyme mix, and pre-mixing ion buffer, dNTPs, PCR stabilizer and enhancer into MasterMix, so that the multi-component mixing steps can be simplified.

● High efficiency: The excellent reverse transcriptase and DNA polymerase ensure high reaction efficiency

● Good reversibility: The polymerase can read through the RNA template with high GC content and complex secondary structure

● Wide applications: High applicability to RNA templates with impurities from different species

● High sensitivity: Templates as low as 1 ng can be accurately identified, especially for low abundance templates

●Example for  RNA Virus Detection●

The nucleic acid of H5 avian influenza was extracted by TGuide S32 Magnetic Viral DNA/RNA Kit (DP604). The FastKing One Step RT-qPCR Kit (Probe) (FP314) was used for RT-qPCR detection using specific H5 avian influenza primers and probes. ABi7500Fast was used for RT-qPCR detection. The results of H5 avian influenza virus antigen (10-2, 10-3, 10-4, 10-5, 10-6 and 10-7 dilution) from 200 μl samples show high virus nucleic acid extraction yield, which can meet the subsequent needs of reverse transcription, PCR, RT-PCR, real-time PCR, etc. The real-time PCR delivers results with high sensitivity, good repeatability and good gradient dilution linearity. Different concentrations of viral nucleic acid samples can be accurately detected.

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