Performance Evaluation of the PowerChek SARS-CoV-2, Influenza A & B Multiplex Real-Time PCR Kit in Comparison with the BioFire Respiratory Panel
Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) and influenza viruses could pose huge challenges to our healthcare system. We evaluated the efficiency of the PowerChek SARS-CoV-2, Influenza A & B Multiplex Real-time PCR Kit (PowerChek; Kogene Biotech, Seoul, Korea) in comparability with the BioFire Respiratory Panels 2 and a couple of.1 (RP2 and RP2.1; bioMérieux, Marcy l’Étoile, France), utilizing 147 nasopharyngeal swabs.
The restrict of detection (LOD) of the PowerChek assay was decided utilizing SARS-CoV-2, influenza A, and B RNA requirements. The LOD values of the PowerChek assay for SARS-CoV-2 and influenza A and B have been 1.12, 1.24, and 0.61 copies/μL, respectively.
The constructive and damaging p.c agreements of the PowerChek assay in contrast with RP2 and RP2.1 have been 97.5% (39/40) and 100% (107/107) for SARS-CoV-2; 100% (39/39) and 100% (108/108) for influenza A; and 100% (35/35) and 100% (112/112) for influenza B, respectively. The efficiency of the PowerChek assay was corresponding to that of RP2 and RP2.1 for detecting SARS-CoV-2 and influenza A and B, suggesting its use in diagnosing SARS-CoV-2 and influenza infections.
Development and Evaluation of AccuPower COVID-19 Multiplex Real-Time RT-PCR Kit and AccuPower SARS-CoV-2 Multiplex Real-Time RT-PCR Kit for SARS-CoV-2 Detection in Sputum, NPS/OPS, Saliva and Pooled Samples
- Rapid and correct detection of the extreme acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is important for the profitable management of the present world COVID-19 pandemic. The real-time reverse transcription polymerase chain response (Real-time RT-PCR) is the most generally used detection approach.
- This analysis describes the growth of two novel multiplex real-time RT-PCR kits, AccuPower® COVID-19 Multiplex Real-Time RT-PCR Kit (NCVM) particularly designed to be used with the ExiStation™48 system (comprised of ExiPrep™48 Dx and Exicycler™96 by BIONEER, Korea) for pattern RNA extraction and PCR detection, and AccuPower® SARS-CoV-2 Multiplex Real-Time RT-PCR Kit (SCVM) designed to be appropriate with producers’ on-market PCR devices.
- The restrict of detection (LoD) of NCVM was 120 copies/mL and the LoD of the SCVM was 2 copies/μL for each the Pan-sarbecovirus gene and the SARS-CoV-2 gene. The AccuPower® kits demonstrated excessive precision with no cross reactivity to different respiratory-related microorganisms.
- The scientific efficiency of AccuPower® kits was evaluated utilizing the following scientific samples: sputum and nasopharyngeal/oropharyngeal swab (NPS/OPS) samples.
- Overall settlement of the AccuPower® kits with a Food and Drug Administration (FDA) accredited emergency use licensed business package (STANDARD™ M nCoV Real-Time Detection package, SD BIOSENSOR, Korea) was above 95% (Cohen’s kappa coefficient ≥ 0.95), with a sensitivity of over 95%.
- The NPS/OPS specimen pooling experiment was carried out to confirm the usability of AccuPower® kits on pooled samples and the outcomes confirmed better than 90% settlement with particular person NPS/OPS samples.
- The scientific efficiency of AccuPower® kits with saliva samples was additionally in contrast with NPS/OPS samples and demonstrated over 95% settlement (Cohen’s kappa coefficient > 0.95). This examine reveals the BIONEER NCVM and SCVM assays are comparable with the present commonplace affirmation assay and are appropriate for efficient scientific administration and management of SARS-CoV-2.
Could pooled samples methodology have an effect on SARS-CoV-2 analysis accuracy utilizing BGI and Sansure-Biotech RT-PCR kits used in Gabon, Central Africa?
- This examine goals at establishing specimens pooling method for the detection of SARS-CoV-2 utilizing the RT-PCR BGI and Sansure-Biotech kits used in Gabon.
- To validate this method, 14 constructive samples, saved at -20°C for 3 to 5 weeks have been analyzed individually (as gold commonplace) and in swimming pools of 5, eight and ten in the identical plate.
- We created 14 swimming pools of 5, Eight and 10 samples utilizing 40 μL from every of the chosen constructive samples blended with 4, 7 and 9 confirmed damaging counterparts in a complete quantity of 200 μL, 320 μL and 400 μL for the swimming pools of 5, Eight and 10 respectively. Both particular person and pooled samples testing was carried out in line with the BGI and Sansure-Biotech RT-PCR protocols used at the Professor Daniel Gahouma Laboratory (PDGL).
- Furthermore, the pooling methodology was additionally examined by evaluating outcomes of 470 unselected samples examined in 94 swimming pools and individually. Results of our experiment confirmed that utilizing a BGI single constructive pattern with cycle threshold (Ct) worth of 28.42, confirmed by particular person testing, detection occurred in all the swimming pools.
- On the opposite samples with Ct >31 weren’t detected in swimming pools of 10 and for these samples (Ct worth as excessive as 37.17) their detection was attainable in pool of 8.
- Regarding the Sansure-Biotech package, constructive samples have been detected in all the pool sizes examined, irrespective of their Ct values. The specificity of the pooling methodology was 100% for the BGI and Sansure-Biotech RT-PCR assays.
- The current examine discovered a rise in the Ct values with pool measurement for the BGI and Sansure-Biotech assays. This pattern was statistically vital (Pearson’s r = 0.978; p = 0,022) utilizing the BGI methodology the place the imply Ct values have been 24.04±1.1, 26.74±1.3, 27.91±1.1 and 28.32±1.1 for the particular person, pool of 5, Eight and 10 respectively.
- The testing of the 470 samples confirmed that one of the 94 swimming pools had a constructive take a look at just like the particular person take a look at utilizing the BGI and Sansure-Biotech kits. The saving of time and economizing take a look at reagents by utilizing the pooling methodology have been demonstrated in this examine.
- Ultimately, the pooling methodology might be used for the analysis of SARS-CoV-2 with out modifying the accuracy of outcomes in Gabon. We advocate a most pool measurement of Eight for the BGI package. For the Sansure-Biotech package, a most pool measurement of 10 can be utilized with out affecting its accuracy in comparison with the particular person testing.
COMPARISON OF COVID-19 LABORATORY DIAGNOSIS BY COMMERCIAL KITS: EFFECTIVITY OF RT-PCR TO THE RT-LAMP.
Coronavirus illness 2019 or COVID-19 attributable to Novel Corona virus/Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2 or 2019-nCoV) is an ongoing pandemic that has rising world results that requires a fast and dependable diagnostic testing.
Quantitative reverse transcription PCR (q-RT-PCR) is the gold commonplace methodology for SARS-CoV- 2 detection. On the different hand, new approaches facilitate the analysis difficulties progressively.
Reverse Transcription Loop-Mediated Isothermal Amplification (RT- LAMP) as one of these novel approaches may contribute to the quicker and cheaper field-based testing on the subject.
The current examine was designed to guage this fast screening diagnostic take a look at that may give outcomes in 30-45 minutes and to match the effectiveness of LAMP to the q-RT-PCR.
The thirty randomly chosen affected person samples have been generated by nasopharyngeal swabs with a portion of the SARS-CoV-2 nucleic sequence.
The pattern of quantification cycle (Cq) values have been examined utilizing RT-LAMP in addition to by typical q-RT-PCR. The affected person samples have been examined with 4 completely different kits (SENSObiz COVID-19 (SARS-CoV-2) LAMP Assay, The QIAseq DIRECT SARS-CoV-2 Kit, Biospeedy SARS-CoV-2 Variant Plus Kit, and CoVirion-CV19-2 SARS-CoV-2 OneStep RT-PCR Kit) and two completely different PCR gadgets (GDS Rotor-Gene Q Thermocycler and Inovia Technologies GenX sequence).
Based on 30 affected person samples, the constructive/damaging ratio (P/N) was 30/Zero as Biospeedy and Covirion (positivity 100%), 28/2 as Qiagen package (pozitify 93.3%) for the samples studied on the Inovia machine whereas the identical samples on the Rotor-Gene machine was 30/Zero as Biospeedy and Covirion (positivity 100%), 29/1 as Qiagen package at the first day (96.7%).
On the fifth day, the samples have been studied in the Inovia machine and respective outcomes have been obtained: 27/Three as Biospeedy (positivity 90%), 16/14 as Qiagen (positivity 53.3%), 28/2 as Covirion package (positivity 93.3%).
When these samples have been studied in the Rotor-Gene machine, it was 29/1 in Biospeedy and Covirion (positivity 96.7%), 19/11 in Qiagen package (positivity 63.3%).
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When these samples have been in contrast with the LAMP methodology it was discovered to be 19/11 (positivity 63.3%) on the first day and 18/12 (positivity %60) on the fifth day. SARS-CoV-2 take a look at research will contribute to the proactive method of growth of fast analysis methods.
LAMP method presents promising outcomes to watch uncovered people and likewise improves screening efforts in potential ports of entry. This article is protected by copyright. All rights reserved.