Nasopharyngeal swabs were taken from all specimens for Ct value analysis. Detection was performed by RT-PCR with the SARS-CoV-2 nucleic acid detection kit (DaAn Gene Co., Ltd.). Real-time PCR was performed on a LightCycler 480II instrument (Roche, Basel, Switzerland). Values below 5 × 102 copies /ml were considered negative.
Sars cov 2 nucleic acid detection kit
Manufactured by Daan Gene
Sourced in China
The SARS-CoV-2 nucleic acid detection kit is a laboratory equipment product designed for the detection of SARS-CoV-2 genetic material. It is used to identify the presence of the SARS-CoV-2 virus in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Powerup sybr green master mix, by Thermo Fisher Scientific (2 mentions)
Trizol reagent, by Thermo Fisher Scientific (2 mentions)
Lightcycler 480 2, by Roche (1 mentions)
Abi quantstudio 5, by Thermo Fisher Scientific (1 mentions)
Magbead viral dna rna kit, by CWBIO (1 mentions)
Primescript rt master mix, by Takara Bio (1 mentions)
7500 fast real time pcr, by Thermo Fisher Scientific (1 mentions)
Magmax viral rna isolation kit, by Thermo Fisher Scientific (1 mentions)
Dsa 100 drop shape analysis system, by Krüss (1 mentions)
8 protocols using sars cov 2 nucleic acid detection kit
1
SARS-CoV-2 Nucleic Acid Detection by RT-PCR
2
Environmental Surface Sampling for SARS-CoV-2
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Environmental surfaces were sampled using swabs with flocked polyester tips moistened with preservation solution after 1 hour of disinfection. An area not exceeding 100 cm2 was rubbed with a moistened swab. The swab was placed in a 15 mL tube with 1.5 mL of Ringer 1/4 solution for transport.
Laboratory confirmation of the virus was performed using real-time reverse transcription polymerase chain reaction (RT-PCR). The fluorescence PCR instrument was purchased from the TianLong company in China (model: Gentier 96E). The full-automatic nucleic acid extractor was from the TianLong company in China (model: GeneRotex 96). The SARS-Cov-2 nucleic acid detection kit was from the Da’an gene company of Sun Yat-Sen University, Guangzhou, China. Cycle threshold values were used to quantify the viral load in each sample, with lower values indicating a higher viral load. A sample was considered positive when the qRT-PCR Ct value was <40.
Laboratory confirmation of the virus was performed using real-time reverse transcription polymerase chain reaction (RT-PCR). The fluorescence PCR instrument was purchased from the TianLong company in China (model: Gentier 96E). The full-automatic nucleic acid extractor was from the TianLong company in China (model: GeneRotex 96). The SARS-Cov-2 nucleic acid detection kit was from the Da’an gene company of Sun Yat-Sen University, Guangzhou, China. Cycle threshold values were used to quantify the viral load in each sample, with lower values indicating a higher viral load. A sample was considered positive when the qRT-PCR Ct value was <40.
3
SARS-CoV-2 Viral Load and Gene Expression
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For detecting the viral load in the supernatant, SARS-CoV-2 RNA was isolated by Magbead Viral DNA/RNA Kit (CWBIO), and SARS-CoV-2 nucleic acid detection kit (Da an Gene Co., Ltd.) was used to quantify viral RNA. For detecting viral RNA or other gene expression in cells, the total RNA was extracted with TRIzol reagent (Thermo Fisher, USA) and reverse-transcribed into cDNA using PrimeScript RT Master Mix (Takara, RR036A) according to the manufacturer’s instructions. RT-qPCR was performed with PowerUp SYBR Green Master Mix (Thermo Fisher, A25742) on the ABI QuantStudio5 (Applied Biosystems, USA). The relative abundance of target RNA was normalized to the human housekeeping gene actin beta ACTB. The primer sequences for detecting SARS-CoV-2 genome or other genes were shown in Supplementary Material Table S1
4
Characterization of Self-Cleaning Glass Surfaces
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The GL-20 G-II High-Speed Refrigerated Centrifuge that experiment used was from Anting Scientific Instruments (Shanghai, China), TU-1901 dual-beam UV visible photometer was from General Analysis Instrument Co., Ltd (Beijing, China), and the pH was debugged with the pHS-3E pH meter from Leici Instrument Factory (Shanghai, China). The water contact angle (WCA) of different SCGS was measured by the DSA 100 drop-shape analysis system (Krüss GmbH, Germany). MagMAX™ Viral RNA Isolation Kit was purchased from Thermo Fisher Scientific (Guangzhou, China). SARS -CoV-2 Nucleic Acid Detection Kit based on fluorescence quantitative PCR was purchased from Daan Gene (Guangzhou, China) and PCR was performed by 7500 Fast Real-Time PCR purchased from Thermo Fisher Scientific (Guangzhou, China). The WCA of SCGS needs to be vacuum dried at 40 °C for two days before measuring. The smartphone (Redmi k30, China) took the images. The RGB values were obtained from the Color recognition picker an application (APP) downed from the mobile app store. The wd-9403 C ultraviolet analyzer (Guangzhou, China) provided a unified photographic environment.
5
SARS-CoV-2 Nucleic Acid Detection Protocol
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Nucleic acid extraction was performed after oropharyngeal swabs specimens were collected from truck drivers according to the “Technical Guidelines for Laboratory Testing of Pneumonia Infected with SARS‐CoV‐2” issued by the National Health Commission of the people's Republic of China. Then, rapid nucleic acid detection was performed on AGS8830 instrument according to the instructions of the SARS‐CoV‐2 nucleic acid detection Kit (Daan Gene Co., Ltd). Briefly, the supporting kit is a single tube and single copy packaging specification, and there is no need to prepare the reaction system. The reagent can be transferred to the amplification detection area after instantaneous centrifugation for 15 s, shaking and mixing for 10 s, and then instantaneous centrifugation for 15 s. Amplification procedure: 50°C 2 min → 95°C 2 min → (95°C 5 s, 60°C 10 s 10 cycles) → (95°C 5 s, 60°C 10 s 32 cycles).
6
SARS-CoV-2 RT-PCR Diagnostic Assay
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Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) laboratory test assays were based on the recommendations of the World Health Organization.[12 ] Throat swab samples were collected from patients with suspected COVID-19 and immediately placed into sterile tubes containing 3 mL of viral transport media. Throat swab RNA was extracted and tested using real-time reverse-transcription polymerase chain reaction (RT-PCR) with SARS-CoV-2 specific primers and probes. Two target genes (ie, open reading frame 1ab and nucleocapsid protein) were simultaneously amplified and tested using real-time RT-PCR. The real-time RT-PCR assay was performed using a SARS-CoV-2 nucleic acid detection kit according to the manufacturer’s instructions (DAAN Gene Co., Ltd., Sun Yat-Sen University, Guangzhou, China), as previously described.[13 (link)]
7
SARS-CoV-2 Detection by RT-PCR
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The methods of laboratory confirmation of SARS-CoV-2 infection have been described (5 (link), 31 (link)). Briefly, SARS-CoV-2 RNA was extracted from throat or nasal swab samples from suspected patients. The presence of SARS-CoV-2 was detected by real-time RT-PCR assay at the local centres of the Chinese Center for Disease Control and Prevention and Tongji Hospital. The SARS-CoV-2 nucleic acid detection kit was used according to the manufacturer’s protocol (DAAN Gene Co., Ltd., of Sun Yat-sen University). These diagnostic methods and criteria were based on the recommendations of the National Institute for Viral Disease Control and Prevention of China.
8
On-chip RT-RPA Assays for COVID-19 Diagnosis
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The diagnostic performance of the on-chip RT-RPA assays in clinical applications was verified with suspected COVID-19 cases tested at Daping Hospital of the Army Medical University. RNA was extracted by an automated nucleic acid workstation with a compatible nucleic acid extraction kit (TianLong Science and Technology Co., Ltd., China) and determined by real-time RT-PCR using a commercial SARS-CoV-2 nucleic acid detection kit (Daan Gene Co., Ltd., China). For RT-PCR, positive samples were those with a Ct value ≤35,whereas negative samples had a Ct value >35. A total of 20 positive samples and 13 negative samples were selected for testing. The RT-RPA assays' sensitivity (i.e., positive coincidence rate) and specificity (i.e., negative coincidence rate) were calculated in comparison to the RT-PCR as a standard method. A Kappa test was used to determine a 95% confidence interval and Kappa value for the coincidence rates.
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