Help differentiate with 40 other viruses
Because of using 3 target genes, only one Realtime cycle is used compared to 2 Realtime cycles according to WHO standards.
One-step RT-PCR COVID-19 Kit Thai Duong is a real-time RT-PCR kit intended for qualitative detection of SARS-CoV-2 from specimens of upper and lower respiratory tracts, such as nasopharyngeal and oropharyngeal swabs, sputum, nasopharyngeal wash or aspirate, nasal aspirate, bronchoalveolar lavage, tracheal aspirate, as recommended by CDC. Test results should be interpreted in conjunction with other testings and clinical evaluations.
One-step RT-PCR COVID-19 Kit Thai Duong enables simultaneous detection of three targets in N gene, two of which specific to SARS-CoV-2; human housekeeping RP gene is used as internal control. The kit contains all reagents necessary for one-step reverse-transcription and amplification using target-specific primers and Taqman probes.
Specimens should be stored at 2-8°C for up to 72 hours after collection. If a delay in extraction is expected, store specimens at -20°C or lower temperature. Extracted nucleic acids should be stored at -20°C
- This product should be used only by personnel trained in real-time PCR techniques and molecular diagnostics.
- All specimens and related materials must be treated as hazardous items and should be handled in a biosafety cabinet in accordance with appropriate biosafety practices.
- Wear disposable gloves, laboratory coat and eye protection during manipulation.
- Clean working area with sodium hypochlorite 0.5%, UV exposure 15 minutes before and after work.
- Do not mix reagents from different kit lots.
- Do not use product after expiration date.
- Do not repeat freeze-thaw of kit components over 5 times. Repeated freezing and thawing may reduce product sensitivity.
SARS-CoV-2, a single-stranded, positive-sense RNA virus, was identified in late December 2019 in Wuhan city. COVID-19, the disease caused by SARS-CoV-2 is now present in more than 190 countries and is declared pandemic by WHO on February 2020. The virus is capable of person-to-person transmission, even from asymptomatic individuals. Seven coronavirus species are known to infect humans. Four of them cause common cold. But infection by the other three including MERS-CoV, SARS-CoV and SARS-CoV-2 result in severe respiratory diseases that could be fatal.
One-step RT-PCR COVID-19 Kit Thai Duong components should be stored in the dark at -20°C. Do not use frost-free freezers for kit storage since freeze-thaw cycles may cause enzyme degradation.
- Consumables (disposable latex or vinyl gloves, RNA-free filter tips 10 µl, 200 µl, 1000 µl)
- RNA extraction kit
- Micropipettes (10 µl, 100 µl, 1000 µl)
- Spindown centrifuge
- Vortex mixer
- –20°C refrigerator
- Real-time thermal cycler (with 4 channels FAM, HEX, TEX/ROX, Cy5)
Standard RNA extraction kits are compatible with this assay. Commercial extraction kits recommended for SARS-CoV-2 detection include bioMérieux NucliSens® systems, QIAamp® Viral RNA Mini Kit, QIAamp® MinElute Virus Spin Kit hoặc RNeasy® Mini Kit (QIAGEN), EZ1 DSP Virus Kit (QIAGEN), Roche MagNA Pure Compact RNA Isolation Kit, Roche MagNA Pure Compact Nucleic Acid Isolation Kit, Roche MagNA Pure 96 DNA, Viral NA Small Volume Kit và Invitrogen ChargeSwitch® Total RNA Cell Kit.
*Reference: Real-Time RT-PCR Panel for Detection 2019-Novel Coronavirus, Centers for Disease Control and Prevention.
1. Thoroughly thaw all reagents on a cold rack. Vortex briefly and spin down.
2. Prepare PCR Master Mix:
3. Pipetting thoroughly Master Mix tube and spin down. Dispense 20 µl of Master Mix into labelled PCR tubes.
4. Add 5 µl of Template. Spin down.
5. PCR program is set as follows:
- Positive Control tube should be prepared separately from Buffer Mix, Enzyme Mix, Primobe Mix, and RNase free water tubes to avoid contamination.
- Do not vortex Enzyme Mix tube.
- Use adequate PCR tube types for your thermal cycler.
- Prepared PCR reactions should be subjected to amplification within 3 hours.
1. Signal detected in Negative control
- Contamination highly possible (due to cross contamination between samples, contamination present in reagents or laboratory environment)
- Repeat RNA extraction, RT and PCR.
- Use a new kit.
- Perform DNA decontamination practices
2. No signal detected in any channel for all samples including Positive control
- Make sure positive controls and PCR reagents are appropriately stored and not expired.
- Check thermal cycling program.
3. No signal detected in any channel for a particular sample
- RNA samples may be unadequately extracted or deteriorated. Make sure RNA are appropriately stored, using RNase-free consumables, check for RNA extraction and repeat the procedure.
The One Step RT-PCR COVID 19 kit Thai Duong has been clinically tested for Limit of Detection (LoD), Clinical evaluation, Inclusivity, Cross-reactivity, Robustness and has the following results:
- Limit of Detection (LoD): 7.558 (3.670-15.564)
- Clinical evaluation: One-step RT-PCR COVID-19 Kit Thai Duong in clinical specimens is 100% in agreement
with WHO-Berlin protocol.
- Inclusivity: One-step RT-PCR COVID-19 Kit Thai Duong primers and probes display high homology with
publicly available 2019-nCoV sequences on GISAID (15/03/2020).
- Cross-reactivity: One-step RT-PCR COVID-19 Kit Thai Duong primers and probes showed no cross
reactivity tested; No cross-reactivity is observed with other 41 tested targets (in vitro).
- Robustness: One-step RT-PCR COVID-19 Kit Thai Duong was validated on CFX96-BioRad, Stratagene
(Agilent), Rotor Gene Q (QIAGEN), Aria (Agilent) and ABI 7500 (Thermo Fisher).
Dr. Le Quang Hoa
Head of Laboratory of Gene Technology, school of Biotechnology anh Food Technology.
Hanoi University of Science and Technology.
Address: 92 Vinhhung street, Hoangmai distrist, Hanoi City, Vietnam Country
Ms. Van: firstname.lastname@example.org/+ 84916451269
Mr. Son: email@example.com/+084943335511
Mon - Sat: 8.am to 4h30.pm