Quick assessment of influenza a virus infectivity with a long-range reverse-transcription quantitative polymerase chain reaction assay
Background: polymerase chain reaction (PCR) which is used to detect viral pathogens because of the high sensitivity and specificity. However, the conventional PCR method can not determine the virus infectivity. Virus infectivity is conventionally checked by methods such as plaque test, although the test like this takes a few days. Long-Range quantitative reverse-transcription PCR (RT-qPCR) has previously been suggested for a rapid assessment of RNA virus infectivity which the loss of infectivity due to the fragmentation of the genome.
Methods: IAV is irradiated with ultraviolet rays of 253.7 nm (UV) to induce genomic strand breaks were confirmed by the full-length RT-PCR assay. The IAV is then subjected to the test plaque, conventional RT-qPCR and RT-qPCR long term to examine the relationship between infectious titer and number of copies. A simple linear regression analysis was conducted to test the correlation between the test results.
Results: A remote RT-qPCR assay was developed and validated for influenza A virus (IAV). Although only a few minutes of UV radiation required to completely disable IAV, genomic RNA remains detectable by conventional RT-qPCR and full-length RT-PCR for the following NS inactivation of the viral genome. A long-distance RT-qPCR assay was then designed using RT-priming at the 3 ‘termini of each segment and subsequent genomic qPCR of the 5’ region. UV-mediated inactivation IAV successfully analyzed by RT-qPCR assay distance especially when targeting the PA of the viral genome. It is also supported by a regression analysis that within RT-qPCR is highly correlated with plaque test (adjusted R2 = 0.931, P = 0.000066).
Conclusion: This study shows that IAV infectivity can be predicted without testing infectivity. Rapid detection of pathogenic IAV has, therefore, been achieved with this sensing technology.

The one-step immunoassay without washing steps for the detection of influenza A virus using isfet
A one-step immunoassay for the detection of influenza A virus were developed using two different microbeads and bottle filter-inserted. Two types of beads with a diameter of 15 (capture beads) and 3 (detection beads) um prepared specifically detect the influenza virus A. Anti-influenza A virus antibody coated on both types of beads, while urease was immobilized only on the detection beads.
Samples of influenza A-positive can form a sandwich complex with the capture and detection of beads; This complex will not pass through the filter, which has a controlled pore size. As the beads used at a concentration detection limit, it will be prevented from crossing the filter; thus, the more it will react with the substrate urea and consequently increase the pH. Influenza A-negative samples will fail to form a sandwich complex in the presence of the capture and detection beads.
Thus, the detection beads will pass through the filter into urea buffer and increasing the pH. Changes in pH in the reaction of urease can be quantitatively measured by indicators such as phenol red or using ion-selective field-effect transistors (isfet). one-step immunoassay is used to detect influenza A viruses in real samples.
Gryphon Ecotropic Packaging Cell Line for Retrovirus (1 x 106 cells, Culture) |
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ABP-RVC-10002C | Allele Biotech | 1 flask | Ask for price |
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LoxP Reporter Cell Lines: LoxP-RFP Human Fibroblast (1 x 106 cells, Culture) |
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ABP-RP-CRFPLOXC | Allele Biotech | 1 flask | Ask for price |
Gryphon Amphotropic Packaging Cell Line for Retrovirus (1 x 106 cells, Culture) |
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ABP-RVC-10001C | Allele Biotech | 1 flask | Ask for price |
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ExoStep Cell Culture |
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ExoS-25-C9 | ImmunoStep | 25 test | EUR 660.6 |
Greiner culture flasks tissue culture treated |
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C6231-200EA | Scientific Laboratory Supplies | PK200 | EUR 231.6 |
Hepes Cell Culture Grade |
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AK3268-0250 | Akron Biotech | 250g | Ask for price |
Hepes Cell Culture Grade |
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AK3268-0500 | Akron Biotech | 500g | Ask for price |
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Water, Cell Culture Grade |
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CA018-600 | GenDepot | 6x1000ml | EUR 148.8 |
EZ-LINE Cell culture Dishes Tissue culture treated, 60X15mm, 500 per case |
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SP31171 | Bio Basic | 1CS, 500UNIT | EUR 230.89 |
EZ-LINE Cell culture Dishes Tissue culture treated, 90x20mm, 200 per case |
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SP31212 | Bio Basic | 1CS, 200UNIT | EUR 162.61 |
ROCKER CELL CULTURE BAG, 2L |
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91-200-80 | CORNING | 1/pk | EUR 450 |
Description: Flexible Containers; Bioprocess Bags and Parts |
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91-200-78 | CORNING | 1/pk | EUR 493.2 |
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91-200-79 | CORNING | 1/pk | EUR 446.4 |
Description: Flexible Containers; Bioprocess Bags and Parts |
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91-200-81 | CORNING | 1/pk | EUR 570 |
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Culture flask CELLine CL 350 |
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DD150102B | Scientific Laboratory Supplies | PK5 | EUR 2312.4 |
Mouse ES Cell Culture Media |
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M16101 | Neuromics | 500 mL | EUR 1336.8 |
6-well Cell Culture Plates |
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abx090688-50plates | Abbexa | 50 plates | EUR 571.2 |
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abx090688-6plates | Abbexa | 6 plates | EUR 142.8 |
Gibco CHO Cell Culture Medium |
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220253 | Scientific Laboratory Supplies | EACH | EUR 57.32 |
Ethanolamine, Cell Culture tested |
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CA071-010 | GenDepot | 100ml | EUR 116.4 |
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Human CD34+ Cell Culture Medium |
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3D Cell Culture Matrix BME Kit |
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K518-100 | Biovision | each | EUR 744 |
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BD Biocoat 10cm cell culture dish |
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354469 | Scientific Laboratory Supplies | PK10 | EUR 112.8 |
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W9200-045 | GenDepot | 450ml | EUR 96 |
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C-01-01-EX | Sceti | 12 sheets | Ask for price |
Description: The CardioDISC (Scaffold for cell culture) is available in Europe and for worldwide shipping via Gentaur. |
culture flask 250ml |
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FLA4286 | Scientific Laboratory Supplies | PK10 | EUR 290.4 |
culture flask 500ml |
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FLA4288 | Scientific Laboratory Supplies | PK5 | EUR 144 |
culture flask 1000ml |
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culture flask 2000ml |
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SP31146 | Bio Basic | 1CS, 500UNIT | EUR 208.27 |
Multifuge X4 Pro cell culture package |
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75016066 | Scientific Laboratory Supplies | EACH | EUR 21742.8 |
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FYG100-300P | Yeastern Biotech | 300 Preps | Ask for price |
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Gelatin Solution , 0.1%, Cell culture tested |
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CA087-100 | GenDepot | 1L | EUR 116.4 |
Pluronic® F-127 *Cell culture tested * |
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20050 | AAT Bioquest | 10 g | EUR 86 |
3D Cell Culture Matrix Duo-Matrix Kit |
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K519-100 | Biovision | each | EUR 718.8 |
40X9mm Cell Culture Plastic Dishes 20/PK |
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DS001-40 | Bio Basic | 1PK, 20UNIT | EUR 64.18 |
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20050-10g | AAT Bioquest | 10 g | EUR 86 |
Description: Pluronic® F-127 is a nonionic surfactant that is 100% active and relatively non-toxic to cells, and frequently used with dye AM esters to improve their water solubility. |
Ampicillin sodium salt, cell culture grade |
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GA0283-100G | Glentham Life Sciences | 100 g | EUR 265.2 |
Ampicillin sodium salt, cell culture grade |
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Ampicillin sodium salt, cell culture grade |
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GA0283-100 | Glentham Life Sciences | 100 | EUR 189.8 |
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GA0283-25 | Glentham Life Sciences | 25 | EUR 71.1 |
Ampicillin sodium salt, cell culture grade |
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Corning CellBIND Surface 35mm Culture Dish |
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3294 | Scientific Laboratory Supplies | PK210 | EUR 261.6 |
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3295 | Scientific Laboratory Supplies | PK126 | EUR 261.6 |
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3296 | Scientific Laboratory Supplies | PK40 | EUR 114 |
60X12mm Cell Culture Plastic Dishes 20/PK |
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DS001-60 | Bio Basic | 1PK, 20UNIT | EUR 64.7 |
90X15mm Cell Culture Plastic Dishes 10/PK |
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DS001-90 | Bio Basic | 1PK, 10UNIT | EUR 65.02 |
ExoStepTM Culture + Standard |
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ExoS-25-CST9 | ImmunoStep | 25 test | EUR 871.2 |
Mammalian Mitochondria Isolation Kit for Cultured Cells |
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20-abx098856 | Abbexa |
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Ceraplex, Premium Cell Culture Suplement(10X) |
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C3318-010 | GenDepot | 100ml | Ask for price |
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C3318-050 | GenDepot | 500ml | Ask for price |
Mitochondria Isolation Kit for Tissue and Cultured Cells |
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KC010100 | Biochain | 1 kit | EUR 310 |
Pluronic F-68, 10% Sol, Cell Culture tested |
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CA072-010 | GenDepot | 100ml | EUR 105.6 |
Techne 1L Culture Vessel |
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TIS4120 | Scientific Laboratory Supplies | EACH | EUR 812.4 |
TPP Culture dish 40x11mm |
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TIS7100 | Scientific Laboratory Supplies | PK900 | EUR 637.2 |
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TIS7106 | Scientific Laboratory Supplies | PK100 | EUR 319.2 |
Water, GlenCell™, suitable for cell culture |
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GK3940-1L | Glentham Life Sciences | 1 l | EUR 79.2 |
ExoFACS? Kit for Cell Culture Media Exosomes |
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K1236-20 | Biovision | each | EUR 973.2 |
HEPES, 99.5%, GlenCell, suitable for cell culture |
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GM5581-1KG | Glentham Life Sciences | 1 kg | EUR 370.8 |
Water, GlenCell™, suitable for cell culture |
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GK3940-1 | Glentham Life Sciences | 1 | EUR 55.4 |
Probenecid *Cell culture tested* *CAS 57-66-9* |
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20060 | AAT Bioquest | 10x72 mg | EUR 86 |
Probenecid *Cell culture tested* *CAS 57-66-9* |
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20060-10x72mg | AAT Bioquest | 10x72 mg | EUR 86 |
Description: Probenecid is an inhibitor of organic-anion transporters located in cell membranes. |
3D Cell Culture Gel 2ml x(soft+medium+hard) |
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P720SMH-6 | 101Bio | - | Ask for price |
3D Cell Culture Matrix Alginate Hydrogel Kit |
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K517-100 | Biovision | each | EUR 666 |
Exosome Isolation kit (for cell culture media) |
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P100 | 101Bio | - | Ask for price |
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P100L | 101Bio | - | Ask for price |
Exosome Isolation kit (for cell culture media) |
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P100S | 101Bio | - | Ask for price |
Tube Culture 26 X 100mm 1636/42 |
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T82050 | Scientific Laboratory Supplies | PK40 | EUR 690 |
Techne 125ml Culture Vessel |
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TIS4090 | Scientific Laboratory Supplies | EACH | EUR 508.8 |
Techne 250ml Culture Vessel |
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TIS4100 | Scientific Laboratory Supplies | EACH | EUR 508.8 |
AnaPrep Cultured Cell DNA Extraction Kit (48) For extracting genomic DNA from up to 5x105 cultured cells |
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Z1322005 | Biochain | 1 kit (48 extractions) Including all required plastic disposables | EUR 216 |
CMRL 1066, CIT CULTURE MEDIA |
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98-304-CV | CORNING | 500 mL/pk | EUR 78 |
Description: Specialty Media; Islet products |
CORNING® ENDOTHELIAL CELL CULTURE MEDIUM, KIT |
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355054 | CORNING | 1/pk | EUR 478.8 |
Description: Advanced Cells - DL; Adv Cell Misc Custom - DL |
L-Alanine, GlenCell, suitable for cell culture |
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GM0361-1KG | Glentham Life Sciences | 1 kg | EUR 303.6 |
PetakaG3 HOT Culture Devices |
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Petaka-HOT | Neuromics | 300 Petaka Devices | EUR 12196.8 |
PetakaG3 LOT Culture Devices |
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Petaka-LOT | Neuromics | 300 Petaka Devices | EUR 12196.8 |
HEPES, 99.5%, GlenCell™, suitable for cell culture |
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GM5581-1 | Glentham Life Sciences | 1 | EUR 276.9 |
L-Arginine, GlenCell, suitable for cell culture |
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GM1092-100G | Glentham Life Sciences | 100 g | EUR 69.6 |
L-Arginine, GlenCell, suitable for cell culture |
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GM1092-1KG | Glentham Life Sciences | 1 kg | EUR 204 |
L-Arginine, GlenCell, suitable for cell culture |
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GM1092-250G | Glentham Life Sciences | 250 g | EUR 93.6 |
L-Arginine, GlenCell, suitable for cell culture |
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GM1092-500G | Glentham Life Sciences | 500 g | EUR 132 |
Suspension Culture Flask 25cm |
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169900 | Scientific Laboratory Supplies | PK200 | EUR 321.6 |
PetakaG3 FLAT Culture Devices |
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Petaka-FLAT | Neuromics | 300 Petaka Devices | EUR 12196.8 |
3D Cell Culture Non-Enzymatic Cell Harvesting Kit |
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K982-100 | Biovision | each | EUR 326.4 |
Tranfferrin, Human Recombinant, Cell culture tested |
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T4600-100 | GenDepot | 100mg | EUR 303.6 |
Tranfferrin, Human Recombinant, Cell culture tested |
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T4600-101 | GenDepot | 1g | EUR 1226.4 |
Tranfferrin, Human Recombinant, Cell culture tested |
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T4600-110 | GenDepot | 10g | EUR 9456 |
N-Acetyl-L-cysteine (>99%, cell culture grade) |
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NALC15-1 | Alpha Diagnostics | 1 g | EUR 270 |
N-Acetyl-L-cysteine (>99%, cell culture grade) |
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NALC15-5 | Alpha Diagnostics | 5 g | EUR 489.6 |
Culture Vessel Fv 1Ltr 1000ml |
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QFV1L | Scientific Laboratory Supplies | EACH | EUR 129.6 |
Culture Vessel Fv 2Ltr 2000ml |
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QFV2L | Scientific Laboratory Supplies | EACH | EUR 205.2 |
SL4R Plus Centrifuge Cell Culture TX1000 Package |
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75016075 | Scientific Laboratory Supplies | EACH | EUR 18357.6 |
L-Glutamine, GlenCell, suitable for cell culture |
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GM8887-1KG | Glentham Life Sciences | 1 kg | EUR 294 |
L-Glutamine, GlenCell, suitable for cell culture |
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GM8887-500G | Glentham Life Sciences | 500 g | EUR 170.4 |
Mammalian Mitochondria Isolation Kit For Tissue & Cultured Cells |
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K288-50 | Biovision | each | EUR 502.8 |
L-Valine, GlenCell™, suitable for cell culture |
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GM3894-1KG | Glentham Life Sciences | 1 kg | EUR 313.2 |
L-Serine, GlenCell™, suitable for cell culture |
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GM7000-1KG | Glentham Life Sciences | 1 kg | EUR 572.4 |
L-Valine, GlenCell™, suitable for cell culture |
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GM3894-1 | Glentham Life Sciences | 1 | EUR 229.3 |
L-Serine, GlenCell™, suitable for cell culture |
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GM7000-1 | Glentham Life Sciences | 1 | EUR 442.9 |
L-Serine, GlenCell™, suitable for cell culture |
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GM7000-100 | Glentham Life Sciences | 100 | EUR 88.6 |
Falcon Culture Slide 4 Well |
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354114 | Scientific Laboratory Supplies | PK24 | EUR 238.8 |
Falcon Culture Slide 8 Well |
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354118 | Scientific Laboratory Supplies | PK24 | EUR 242.4 |
Duran 1L Culture erlen. Flask |
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FLA42780 | Scientific Laboratory Supplies | PK10 | EUR 369.6 |
Culture Tube 13x100mm 8ml PS |
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T4004 | Scientific Laboratory Supplies | PK1000 | EUR 121.2 |
Tissue Culture Plate, 12 Well |
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TCP20-12 | Bio Basic | 1 UNIT | EUR 64.18 |
Tissue Culture Plate, 48 Well |
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TCP20-48 | Bio Basic | 1 UNIT | EUR 64.18 |
Tissue Culture Plate, 6 Well |
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TCP20-6 | Bio Basic | 1 UNIT | EUR 64.18 |
Tissue Culture Plate, 96 Well |
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TCP20-96 | Bio Basic | 1 UNIT | EUR 64.18 |
Culture Tube PS 12x75mm Bulk |
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TIS5400 | Scientific Laboratory Supplies | PK500 | EUR 194.4 |
Culture Tube PP 12x75mm Bulk |
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TIS5402 | Scientific Laboratory Supplies | PK500 | EUR 222 |
Bovine Collagen Type I (Skin, >97%) for cell culture |
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CLGN16-N-1 | Alpha Diagnostics | 5 mg | EUR 634.8 |
Insect Cell Medium: TNM-FH Insect Culture Medium |
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ABP-MED-10001 | Allele Biotech | 1 liter | Ask for price |
Exosome Isolation kit (for stem cell culture media) |
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P107S | 101Bio | - | Ask for price |
Exosome Isolation kit (for stem cell culture media) |
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P107 | 101Bio | - | Ask for price |
L-Leucine, GlenCell™, suitable for cell culture |
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GM0001-1KG | Glentham Life Sciences | 1 kg | EUR 250.8 |
L-Proline, GlenCell™, suitable for cell culture |
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GM9897-1KG | Glentham Life Sciences | 1 kg | EUR 361.2 |
L-Leucine, GlenCell™, suitable for cell culture |
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GM0001-1 | Glentham Life Sciences | 1 | EUR 177.9 |
L-Alanine, GlenCell™, suitable for cell culture |
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GM0361-1 | Glentham Life Sciences | 1 | EUR 221.5 |
L-Proline, GlenCell™, suitable for cell culture |
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GM9897-1 | Glentham Life Sciences | 1 | EUR 268.9 |
L-Proline, GlenCell™, suitable for cell culture |
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GM9897-100 | Glentham Life Sciences | 100 | EUR 63.3 |
3D Cell Culture HTS Cell Viability Complete Assay Kit |
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K948-100 | Biovision | each | EUR 411.6 |
Tube Culture With Svl Plastic |
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B99485 | Scientific Laboratory Supplies | PK40 | EUR 442.8 |
Culture Tube PP 17x100mm Bulk |
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TIS5406 | Scientific Laboratory Supplies | PK500 | EUR 336 |
L-Arginine, GlenCell™, suitable for cell culture |
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GM1092-1 | Glentham Life Sciences | 1 | EUR 138.5 |
L-Arginine, GlenCell™, suitable for cell culture |
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GM1092-100 | Glentham Life Sciences | 100 | EUR 27.8 |
L-Arginine, GlenCell™, suitable for cell culture |
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GM1092-250 | Glentham Life Sciences | 250 | EUR 47.4 |
L-Arginine, GlenCell™, suitable for cell culture |
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GM1092-500 | Glentham Life Sciences | 500 | EUR 79.1 |
Duran 250ml Culture erlen. Flask |
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FLA42720 | Scientific Laboratory Supplies | PK10 | EUR 288 |
Duran 300ml Culture erlen. Flask |
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FLA42740 | Scientific Laboratory Supplies | PK10 | EUR 144 |
Duran 500ml Culture erlen. Flask |
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FLA42760 | Scientific Laboratory Supplies | PK10 | EUR 178.8 |
Culture Tube 125x16mm Soda Lime |
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TES6110 | Scientific Laboratory Supplies | PK486 | EUR 51.31 |
Exosome Purification and Detection Kit (Cell Culture) |
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abx290025-25tests | Abbexa | 25 tests | EUR 777.6 |
Culture Tube 75x10mm Borosilicate |
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TES5120 | Scientific Laboratory Supplies | PK1000 | EUR 62.8 |
Culture Tube 75x12mm Borosilicate |
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TES5122 | Scientific Laboratory Supplies | PK1000 | EUR 61.2 |
Culture Tube 100x13mm Borosilicate |
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TES5124 | Scientific Laboratory Supplies | PK1000 | EUR 91.2 |
Culture Tube 100x16mm Borosilicate |
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TES5128 | Scientific Laboratory Supplies | PK1000 | EUR 124.8 |
Culture Tube 150x16mm Borosilicate |
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TES5132 | Scientific Laboratory Supplies | PK1000 | EUR 171.6 |
Culture Tube 150x18mm Borosilicate |
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TES5135 | Scientific Laboratory Supplies | PK500 | EUR 136.8 |
Culture Tube 150x20mm Borosilicate |
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TES5136 | Scientific Laboratory Supplies | PK500 | EUR 322.8 |
Receiver operating characteristics (ROC) plot analysis shows the value of area under the curve (AUC) of 0.931; the sensitivity and specificity of the test was 80% and 90%, respectively, with a cutoff value of 0.9986. These results indicate that one-step immunoassay can increase the sensitivity of detection of influenza A viruses in real samples.