Using 200 ul of nasopharyngeal swab material, nucleic acid was extracted by the bioMerieux easyMAG automated extractor. Specimens were tested using the xTAG Respiratory Virus Panel (RVP) assay for influenza A (subtype H1 and H3), influenza B, RSV (subtype A and B), parainfluenza (1–4), coronavirus (NL63, OC43, HKU1 and 229E), human metapneumovirus, entero-rhinovirus, and adenovirus as per the manufacturer's protocol (Luminex Molecular Diagnostics, Inc., Toronto, Ontario).
Easymag automated extractor
Manufactured by bioMérieux
Sourced in United States
The EasyMAG automated extractor is a laboratory instrument designed for nucleic acid extraction. It performs automated sample preparation, allowing for efficient and reproducible extraction of DNA and RNA from a variety of sample types. The core function of the EasyMAG is to extract and purify nucleic acids in a streamlined and standardized process.
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Lab products found in correlation
6 protocols using easymag automated extractor
1
Comprehensive Respiratory Virus Screening
2
HHV-8 Detection in B Cells via PCR
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For the in vitro experiments, a total of 5×105 B cells and 500 µl of culture supernatant were collected post-HHV-8 exposure, pelleted, and assayed by PCR (50 (link), 51 (link)). Samples were treated with 1 µl DNase in 10 µl buffer (Sigma) for 15 min and then lysed in easyMAG buffer (NucliSENS). DNA was extracted with an easyMAG automated extractor (bioMérieux). Phocine herpesvirus was added as an internal control (52 (link)). DNA was mixed with a primer set specific for HHV-8 K8.1 (51 (link)), and the real-time PCR was done using a 7000, 7500, or ViiA7 ABI system (Applied Biosystems). B cells isolated from PBMC obtained from KS cases and controls were treated and tested for HHV-8 DNA as described above.
3
Viral RNA Extraction and RT-PCR Detection
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Viral RNA from samples were extracted using the easyMAG® automated extractor (BioMerieux, Quebec, Canada), according to manufacturer’s instructions. The input volumes for plasma and cerebrospinal fluid were 200 µl and the output volume was 55 µl. For urine the corresponding volumes were 200 µl and 110 µl respectively. A one-step RT-PCR was then performed employing TaqMan®Fast VirusOne-Step RT-PCR Master Mix (ABI) according to the protocol of Pabbaraju et al. [20 (link)].
4
Viral Nucleic Acid Extraction Using EasyMag
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Isolation of viral nucleic acid from control material and patient specimens was performed using an EasyMag automated extractor (bioMerieux, Durham, NC, USA) as previously described for the SRT-PCR assay.4 (link)
5
Automated Viral Nucleic Acid Extraction
6
Multiplex Assay for Enterovirus Detection
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High and low viral loads of representative EVs from the different species including CV-A2, CV-A4, CV-A5, CV-A6, CV-A16 and EV-A71 for species A; CV-A9, CV-B2, CV-B3, CV-B4, CV-B5, E-4, E-9, E-18, E-25, E-30, E-82, EV-B86 and EV-B101 from species B; vaccine strains of PV-1, PV-2 and PV-3 from species C and EV-D68 and EV-D70 from species D were tested by the multiplex assay. Human PeVs tested by the assay included HPeV-1 and 3. All EVs were typed based on the partial sequence of the VP2 gene [4] (link), or of VP1 for CV-A5 and CV-B2 [13] (link).
To assess any competitive inhibition of target detection in cases with co-infections of EV and HPeVs, spiked samples were tested with different concentrations of both viruses. Cultured CV-B4 was spiked to give Ct values ranging from 20.17 to 32.47 and HPeV-3 was spiked to give Ct values ranging from 22.85 to 32.64 into universal transport media (UTM) and extracted using the using the easyMAG® automated extractor (BioMérieux, Durham, NC, USA). All extracts were tested by the single and multiplex assays.
To assess any competitive inhibition of target detection in cases with co-infections of EV and HPeVs, spiked samples were tested with different concentrations of both viruses. Cultured CV-B4 was spiked to give Ct values ranging from 20.17 to 32.47 and HPeV-3 was spiked to give Ct values ranging from 22.85 to 32.64 into universal transport media (UTM) and extracted using the using the easyMAG® automated extractor (BioMérieux, Durham, NC, USA). All extracts were tested by the single and multiplex assays.
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