To enrich RT-qPCR testing for analysis of extreme acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infections, many international locations have launched using fast antigen exams. As they typically show decrease real-life performances than anticipated, their right positioning as frontline screening continues to be controversial. Regardless of the shortage of information from day by day medical use, third era microfluidic assays (such because the LumiraDx SARS-CoV-2 Ag take a look at) have just lately been instructed to have comparable performances to RT-qPCR and have been proposed as various diagnostic instruments.
By analyzing 960 nasopharyngeal swabs from 960 topics on the emergency division admissions of a tertiary COVID-19 hospital, LumiraDx assay demonstrated a specificity of 97% (95% CI: 96-98), and a sensitivity of 85% (95% CI: 82-89) as compared with RT-qPCR, which will increase to 91% (95% CI: 86-95) for samples with a cycle threshold ≤ 29. Fifty false-negative LumiraDx-results have been confirmed by direct quantification of genomic SARS-CoV-2 RNA by way of droplet-digital PCR (median (IQR) load = 5880 (1657-41,440) copies/mL). Subgenomic N and E RNAs have been detected in 52% (n = 26) and 56% (n = 28) of them, respectively, supporting the presence of lively viral replication. Total, the LumiraDx take a look at complies with the minimal efficiency necessities of the WHO. But, the danger of a misrecognition of sufferers with lively COVID-19 persists, and the necessity for confirmatory RT-qPCR shouldn’t be amended.
Growth of a Tetraplex qPCR for the Molecular Identification and Quantification of Human Enteric Viruses, NoV and HAV, in Fish Samples
Human enteric viruses resembling norovirus (NoV) and hepatitis A virus (HAV) are a number of the most necessary causes of foodborne infections worldwide. Normally, an infection by way of fish consumption isn’t a priority concerning these viruses, since fish are primarily consumed cooked. Nevertheless, within the final years, uncooked fish consumption has turn into more and more widespread, particularly involving using seabass and gilthead seabream in dishes like sushi, sashimi, poke, and carpaccio.
Due to this fact, the danger for viral an infection by way of the consumption of uncooked fish has additionally elevated. On this research, a virologic screening was carried out in 323 fish specimens captured alongside the Portuguese coast utilizing a tetraplex qPCR optimised for 2 templates (plasmid and in vitro transcribed RNA) to detect and quantify NoV GI, NoV GII and HAV genomes. A distinction of roughly 1-log was discovered between using plasmid or in vitro transcribed RNA for molecular-based quantifications, exhibiting an underestimation of genome copy-number equivalents utilizing plasmid standard-based curves.

Moreover, the presence of NoV genomic RNA in a pool of seabass brains was recognized, which was proven to cluster with a serious group of human norovirus sequences from genogroup I (GI.1) by phylogenetic evaluation. Not one of the analysed fish revealed the presence of NoV GII or HAV. This end result corroborates the speculation that enteric viruses flow into in seawater or that fish have been contaminated throughout their transportation/dealing with, representing a possible threat to people by way of uncooked or undercooked fish consumption.
Screening of Reference Genes for RT-qPCR in Rooster Adipose Tissue and Adipocytes
Reverse transcription quantitative real-time PCR is essentially the most generally used technique to detect gene expression ranges. In experiments, it’s typically essential to right and standardize the expression degree of goal genes with reference genes. Due to this fact, it is rather necessary to pick secure reference genes to acquire correct quantitative outcomes. Though software examples of reference genes in mammals have been reported, no research have investigated using reference genes in finding out the expansion and growth of adipose tissue and the proliferation and differentiation of preadipocytes in chickens.
On this research, GeNorm, a reference gene stability statistical algorithm, was used to research the expression stability of 14 candidate reference genes within the stomach adipose tissue of broilers at 1, 4, and seven weeks of age, the proliferation and differentiation of main preadipocytes, in addition to instantly remoted preadipocytes and mature adipocytes. The outcomes confirmed that the expression of the TATA field binding protein (TBP) and hydroxymethylbilane synthase (HMBS) genes was most secure through the development and growth of stomach adipose tissue of broilers, the expression of the peptidylprolyl isomerase.
A (PPIA) and HMBS genes was most secure through the proliferation of main preadipocytes, the expression of the TBP and RPL13 genes was most secure through the differentiation of main preadipocytes, and the expression of the TBP and HMBS genes was most secure in instantly remoted preadipocytes and mature adipocytes. These outcomes present reference bases for precisely detecting the mRNA expression of useful genes in adipose tissue and adipocytes of chickens.
Stopping Chagas illness: A brand new RT-qPCR technique for fast and particular quantification of viable Trypanosoma cruzi for meals security
With out standardized strategies for quickly detecting in meals matrices viable T. cruzi, foodborne outbreaks stay uncared for. On this work, a reverse-transcriptase real-time PCR (RT-qPCR) mRNA-based approach was developed for the fast and particular detection and quantification of viable Trypanosoma cruzi in açai fruits and juice. The tactic makes use of particular primer concentrating on area on the cyt b gene.
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RB94-YPD100 | T-Pro Biotechnology | 100preps/Kit | EUR 193.2 |
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T-Pro Genomic DNA Mini Kit (100) |
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20-abx098887 | Abbexa |
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T-Pro LumiFast Plus Chemiluminescence Detection Kit (ECL Kit) |
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JT96-K002M | T-Pro Biotechnology | 250ml*2/Kit | EUR 244.8 |
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HiScript II Q Select RT SuperMix for qPCR (+g DNA wiper) |
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R233-01 | Vazyme | 100 rxn (20 μl/rxn) | EUR 292.8 |
T-Pro Endotoxin Removal Plasmid Midi kit (25) |
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RB94-EPI020 | T-Pro Biotechnology | 20preps/Kit | EUR 297.6 |
T-Pro Endotoxin Removal Plasmid Maxi kit (10) |
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RB94-EPM010 | T-Pro Biotechnology | 10preps/Kit | EUR 318 |
T-Pro Plant Genomic DNA Midi Kit (20) |
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RB94-PGM020 | T-Pro Biotechnology | 20preps/kit | EUR 266.4 |
T-Pro Plant Genomic DNA Mini Kit (100) |
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RB94-PGS100 | T-Pro Biotechnology | 100preps/kit | EUR 266.4 |
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T-Pro Transfer Blotting buffer (10X) |
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T-Pro Western Blot Stripping Reagent |
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JT90-R001M | T-Pro Biotechnology | 500ml/BT | EUR 398.4 |
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JT90-R001S | T-Pro Biotechnology | 100ml/BT | EUR 181.2 |
T-Pro EZ stain Gel solution |
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JT90-R005S | T-Pro Biotechnology | 100ml/BT | EUR 151.2 |
T-Pro P-Fect Transfection Reagent |
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JT97-N005M | T-Pro Biotechnology | 1.0ml/vial | EUR 266.4 |
QPCR Kit QPCR Mastermix DNA |
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Library Amplification SuperMix |
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cDNA Synthesis SuperMix |
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Evo? cDNA Supermix |
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Novo? cDNA Supermix |
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M1169-25 | Biovision | EUR 346.8 |
OneScriptcDNA Synthesis SuperMix |
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G451 | ABM | 25 x 20 ul reactions | EUR 106.8 |
OneScriptcDNA Synthesis SuperMix |
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T-Pro BCA Protein Assay Reagent A |
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JB04-D001A | T-Pro Biotechnology | 500ml/BT | EUR 204 |
T-Pro BCA Protein Assay Reagent B |
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JB04-D001B | T-Pro Biotechnology | 12ml/BT | EUR 140.4 |
T-Pro Fast Blocking Buffer (in PBS) |
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JK92-W001 | T-Pro Biotechnology | 500ml/BT | EUR 224.4 |
T-Pro Fast Blocking Buffer (in TBS) |
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T-Pro Laemmli (SDS sample) Reagent (reducing 4X) |
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JB06-F002 | T-Pro Biotechnology | 10ml/BT | EUR 151.2 |
T-Pro Tris-Glycine-SDS running buffer (10X) |
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JB07-G001 | T-Pro Biotechnology | 500ml/BT | EUR 162 |
T-Pro Tris-Glycine-Native running buffer (10X) |
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T-Pro Semi Dry Transfer Blotting buffer (10X) |
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T-Pro Protein Free Blocking Buffer (in PBS) |
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JK92-W003 | T-Pro Biotechnology | 500ml/BT | EUR 224.4 |
T-Pro Protein Free Blocking Buffer (in TBS) |
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JK92-W004 | T-Pro Biotechnology | 500ml/BT | EUR 224.4 |
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T-Pro Nonliposomal Transfection Reagent II (NTR II) |
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FastPfu PCR SuperMix (-dye) |
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OneScriptPlus cDNA Synthesis SuperMix |
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G453 | ABM | 25 x 20 ul reactions | EUR 116.4 |
OneScriptPlus cDNA Synthesis SuperMix |
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G454 | ABM | 100 x 20 ul reactions | EUR 202.8 |
T-Pro Laemmli (SDS sample) Reagent (non-reducing 4X) |
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JB06-F003 | T-Pro Biotechnology | 10ml/BT | EUR 151.2 |
AXYGEN® AXYPET® PRO STARTER KIT: 4 PIPETTORS, PLEXI STAND, SAMPLE TIPS |
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AP-STR-KIT-P | CORNING | 1/pk | EUR 865.2 |
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T-Pro Washing buffer in PBS and Tween-20 (10X) |
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JB09-I003 | T-Pro Biotechnology | 500ml/BT | EUR 172.8 |
T-Pro Washing buffer in TBS and Tween-20 (10X) |
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Frit Kit |
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FRIT-KIT | Next Advance | 1each | EUR 148.8 |
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T-Pro Phosphate-Buffered Saline (PBS, 10X) for western blot washing |
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JB09-I001 | T-Pro Biotechnology | 500ml/BT | EUR 162 |
T-Pro Tris-Buffered Saline (TBS, 10X) for western blot washing |
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Column Packing Kit |
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PACK-KIT | Next Advance | 1pack | EUR 1242 |
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PCR Mycoplasma Detection Kit |
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M034-Kit | TOKU-E | Kit | EUR 319.2 |
PCR SuperMix for PAGE (+dye) |
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20-abx098004 | Abbexa |
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Shikari (S-ATD) Denosumab (Prolia) Qualitative ELISA kit |
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DEN-QLS-PRO | Matriks Biotek | 1x96-wells test plate | EUR 999 |
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QPCR Positive Control Kit |
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M1126-100 | Biovision | EUR 483.6 |
qPCR Control Template Kit |
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Lentivirus qPCR Quantification Kit |
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Entrans qPCR Probe Kit |
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RK21209 | Abclonal | 40 RXN | Ask for price |
QPCR Kit DNA Staphylococ |
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MOL6370 | Scientific Laboratory Supplies | EACH | EUR 839.04 |
QPCR Kit DNA Bifidobacte |
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MOL6418 | Scientific Laboratory Supplies | EACH | EUR 842.46 |
QPCR Kit DNA Clostridium |
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MOL6422 | Scientific Laboratory Supplies | EACH | EUR 876.66 |
QPCR Kit DNA Lactobacill |
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MOL6474 | Scientific Laboratory Supplies | EACH | EUR 876.66 |
QPCR Kit DNA Anacardium |
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MOL6486 | Scientific Laboratory Supplies | EACH | EUR 842.46 |
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MOL6498 | Scientific Laboratory Supplies | EACH | EUR 876.66 |
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MOL6550 | Scientific Laboratory Supplies | EACH | EUR 876.66 |
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MOL6562 | Scientific Laboratory Supplies | EACH | EUR 842.46 |
QPCR Kit DNA Phacochoeru |
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MOL6574 | Scientific Laboratory Supplies | EACH | EUR 876.66 |
QPCR Kit DNA Pleuronecte |
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MOL6582 | Scientific Laboratory Supplies | EACH | EUR 842.46 |
QPCR Kit DNA Pollachius |
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MOL6594 | Scientific Laboratory Supplies | EACH | EUR 842.46 |
QPCR Kit DNA Bifidobacte |
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MOL6614 | Scientific Laboratory Supplies | EACH | EUR 839.04 |
QPCR Kit DNA Clostridium |
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MOL6630 | Scientific Laboratory Supplies | EACH | EUR 772.92 |
QPCR Kit DNA Enterococcu |
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MOL6634 | Scientific Laboratory Supplies | EACH | EUR 839.04 |
QPCR Kit DNA Enterococcu |
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MOL6638 | Scientific Laboratory Supplies | EACH | EUR 804.84 |
QPCR Kit DNA Pseudomonas |
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MOL6666 | Scientific Laboratory Supplies | EACH | EUR 839.04 |
QPCR Kit DNA Cyclospora |
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MOL6694 | Scientific Laboratory Supplies | EACH | EUR 839.04 |
QPCR Kit DNA Campylobact |
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MOL6702 | Scientific Laboratory Supplies | EACH | EUR 839.04 |
QPCR Kit DNA Escherichia |
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MOL6710 | Scientific Laboratory Supplies | EACH | EUR 839.04 |
QPCR Kit DNA Escherichia |
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MOL6718 | Scientific Laboratory Supplies | EACH | EUR 1081.86 |
QPCR Kit DNA Enteropatho |
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MOL6734 | Scientific Laboratory Supplies | EACH | EUR 839.04 |
QPCR Kit DNA Francisella |
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MOL6738 | Scientific Laboratory Supplies | EACH | EUR 839.04 |
QPCR Kit DNA Mycobacteri |
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MOL6750 | Scientific Laboratory Supplies | EACH | EUR 839.04 |
QPCR Kit DNA Shewanella |
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MOL6758 | Scientific Laboratory Supplies | EACH | EUR 772.92 |
QPCR Kit DNA Aggregatiba |
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MOL6762 | Scientific Laboratory Supplies | EACH | EUR 772.92 |
QPCR Kit DNA Acinetobact |
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MOL6766 | Scientific Laboratory Supplies | EACH | EUR 839.04 |
QPCR Kit DNA Ajellomyces |
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MOL6770 | Scientific Laboratory Supplies | EACH | EUR 772.92 |
QPCR Kit DNA Balamuthia |
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MOL6806 | Scientific Laboratory Supplies | EACH | EUR 772.92 |
QPCR Kit DNA Bordetella |
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MOL6818 | Scientific Laboratory Supplies | EACH | EUR 804.84 |
QPCR Kit DNA Clostridium |
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MOL6826 | Scientific Laboratory Supplies | EACH | EUR 772.92 |
QPCR Kit DNA Clostridium |
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MOL6830 | Scientific Laboratory Supplies | EACH | EUR 839.04 |
QPCR Kit DNA Clostridium |
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MOL6838 | Scientific Laboratory Supplies | EACH | EUR 804.84 |
QPCR Kit DNA Cytomegalov |
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MOL6854 | Scientific Laboratory Supplies | EACH | EUR 839.04 |
QPCR Kit DNA Enterococcu |
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MOL6866 | Scientific Laboratory Supplies | EACH | EUR 772.92 |
QPCR Kit DNA Haemophilus |
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MOL6918 | Scientific Laboratory Supplies | EACH | EUR 839.04 |
QPCR Kit RNA Coronavirus |
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MOL6946 | Scientific Laboratory Supplies | EACH | EUR 820.8 |
QPCR Kit DNA Klebsiella |
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MOL8156 | Scientific Laboratory Supplies | EACH | EUR 1220.94 |
QPCR Kit DNA Leishmania |
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MOL8164 | Scientific Laboratory Supplies | EACH | EUR 839.04 |
The utmost restoration price of T. cruzi from inoculated açai juice was 82.50%. The restrict of detection and quantification in açai juice was 10 parasites/mL for RT-qPCR (mRNA-based) and qPCR (DNA-based). The RT-qPCR effectivity was estimated at 97.27% with an R2 of 0.994. The RT-qPCR was proven to have the ability to discriminate between viable and nonviable cells. This technique supplies a great tool for fast evaluation of low concentrations of viable T. cruzi in naturally contaminated meals samples, and may be utilized industrially as a high quality and safety technique.
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