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.

 Quick assessment of influenza a virus infectivity with a long-range reverse-transcription quantitative polymerase chain reaction assay
Quick assessment of influenza a virus infectivity with a long-range reverse-transcription quantitative polymerase chain reaction assay

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.

Cell Culture Grade Water, Cell Culture Tested, Sterile

CCM2081-1000 1000 mL
EUR 82.64

Gryphon Ecotropic Packaging Cell Line for Retrovirus (1 x 106 cells, Culture)

ABP-RVC-10002C 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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Cell Culture Plate

CCP01000-00-CellCulturePlate6Well2CellCulturePlate96Well2 Cell Culture Plate,6 Well*2Cell Culture Plate,96 Well*2
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10X HBSS Cell culture

IBS-BH011 1L
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CRFK CELL TUBE CULTURE

63-0600 each Ask for price

Water for Cell Culture

TBS5050 500ml
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IBS-CB016 1L
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T225 Cell Culture Flask

P0230 25/case
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25-511 6 x 500 mL/Unit
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25-511B 6 x 1000 mL
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IB42010 1L
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IB42011 6x1L
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IB42012 12x1L
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IB42020 2L
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IB42021 6x2L
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IB42030 10L
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EZ-LINE Cell culture Dishes Tissue culture treated, 60X15mm, 500 per case

SP31171 1CS, 500UNIT
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EZ-LINE Cell culture Dishes Tissue culture treated, 90x20mm, 200 per case

SP31212 1CS, 200UNIT
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25 cm2 Cell Culture Flask

CCF00025-00-10Bag 10 / Bag
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25 cm2 Cell Culture Flask

CCF00025-200Case 200 / Case
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75 cm2 Cell Culture Flask

CCF00075-00-5Bag 5 / Bag
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75 cm2 Cell Culture Flask

CCF00075-100Case 100 / Case
EUR 176.04

Human MEP Cell Culture Medium (human megakaryocyte–erythroid progenitor cell culture medium)

P770 8 rxn
EUR 499

ROCKER CELL CULTURE BAG, 2L

91-200-80 1/pk
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175 cm2 Cell Culture Flask

CCF00175-40Case 40 / Case
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225 cm2 Cell Culture Flask

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Cell Culture Plate, 6 Well

CCP01006-50Case 50 / Case
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CCP01096-100Case 100 / Case
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91-200-78 1/pk
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91-200-79 1/pk
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91-200-81 1/pk
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Mouse ES Cell Culture Media

M16101 500 mL
EUR 1336.8

Cellulase, Plant Culture Tested

PCT1518-100KU 1 unit
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6-well Cell Culture Plates

abx090688-50plates 50 plates
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abx090688-6plates 6 plates
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25-105 100 Plates/Unit
EUR 119.75
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HEPES. cell culture tested - kg

PM-B2093/1000 kg
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abx090688-100l 100 µl
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abx090688-200l 200 µl
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25-106 100 Plates/Unit
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25-107 100 Plates/Unit
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25-221 100 plates
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25-281 6 Inserts/Plate, 24/Unit
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25-282 6 Inserts/Plate, 24/Unit
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25-283 6 Inserts/Plate, 24/Unit
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Imiquimod for cell culture, 98%

75054 250 Mg
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PM-B2093/100 100g
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PM-B2093/250 250g
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PM-B2093/500 500g
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MR1015 1 bottle
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Ethanolamine, Cell Culture tested

CA071-010 100ml
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25-285 12 Inserts/Plate, 48/Unit
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25-286 12 Inserts/Plate, 48/Unit
EUR 112.46
Description: 8µm PC Membrane, Sterilized

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25-287 12 Inserts/Plate, 48/Unit
EUR 112.46
Description: 1µm PC Membrane, Sterilized

Cell Culture Inserts, 24-Well

25-288 12 Inserts/Plate, 48/Unit
EUR 112.46
Description: 3µm PC Membrane, Sterilized

Cell Culture Inserts, 24-Well

25-289 12 Inserts/Plate, 48/Unit
EUR 112.46
Description: 8µm PC Membrane, Sterilized

Yeast Extract for cell culture

52184 100 Gms
EUR 9.41
Description: Part D

Yeast Extract for cell culture

52184-1 500 Gms
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Nunc Cell Culture Plate Holder

LBI10241 each
EUR 227.81

ITS Cell Culture Supplement(100×)

ITSS-10201 10mL
EUR 59

Human CD34+ Cell Culture Medium

P771 8 rxn
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HEPES Buffer (Cell culture grade)

IBS-CB023 100 mL
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100 mm Cell Culture Dish Holder

LBI10235 each
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Greiner culture flasks tissue culture treated - PK200

C6231-200EA PK200
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LBI10233 each
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60 mm Cell Culture Dish Holder

LBI10234 each
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3D Cell Culture Matrix BME Kit

K518-100 each
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Cell Culture Freezing Medium (FBS)

cAP-22 50ml
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nBIONIX Hypoxic Cell Culture Kit

nBIONIX-3 1 Kit
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Culture flask CELLine CL 350 - PK5

DD150102B PK5
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W9200-045 450ml
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DMSO Bio-Max, Cell Culture Grade

40470005-1 50 mL
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40470005-2 100 mL
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40470005-3 25 mL
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40470005-4 250 mL
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Mitochondria Isolation Kit (Cultured Cells)

K1138-10 10rxns
EUR 28
Description: Mitochondria Isolation Kit

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K1138-20 20rxns
EUR 44
Description: Mitochondria Isolation Kit

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K1138-30 30rxns
EUR 56
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Cell Culture Supernatant Diluent Kit

DKC100 96 Tests
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CardioDISC (Scaffold for cell culture)

C-01-01-EX 12 sheets Ask for price
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500mL Water Cell Culture Grade - PK6

25-055-CV PK6
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Gibco CHO Cell Culture Medium - EACH

220253 EACH
EUR 64.48

EZ-LINE Cell culture Dishes Tissue culture treated, 35X10mm, PS, Sterile TC treated, 500 per case

SP31146 1CS, 500UNIT
EUR 208.27

ExoStep Culture

ExoS-25-C9 25 test
EUR 423.5
Description: Exosome purification and FACS analysis kit of exosomes derived from cell culture

Mitochondrion Extraction Kit (Cultured Cells)

MBS9718637-200Tests 200Tests
EUR 230

Mitochondrion Extraction Kit (Cultured Cells)

MBS9718637-50Tests 50Tests
EUR 170

Mitochondrion Extraction Kit (Cultured Cells)

MBS9718637-5x200Tests 5x200Tests
EUR 1010

Cell Culture Water Pyrogen free - 100ml

LM-T1707/100 100ml
EUR 2.59

Cell Culture Water Pyrogen free - 1000ml

LM-T1707/1000 1000ml
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Cell Culture Water Pyrogen free - 500ml

LM-T1707/500 500ml
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Pluronic F-127 *Cell culture tested *

MBS8584854-100mg 100mg
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Pluronic F-127 *Cell culture tested *

MBS8584854-1g 1g
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Pluronic F-127 *Cell culture tested *

MBS8584854-5x1g 5x1g
EUR 680

1L Water Cell Culture Grade WFI - PK6

25-055-CM PK6
EUR 95.85

3D Cell Culture Gel, hard (Col-Tgel)

P720H-10 10 mL
EUR 389

3D Cell Culture Gel, hard (Col-Tgel)

P720H-2 2 mL
EUR 189

3D Cell Culture Gel, soft (Col-Tgel)

P720S-10 10 mL
EUR 389

3D Cell Culture Gel, soft (Col-Tgel)

P720S-2 2 mL
EUR 189

i-poly Cell Culture Plate (6 well)

IPY-31006 50 ea/cs
EUR 33
Description: Type: Plate

i-poly Cell Culture Plate (24 well)

IPY-31024 50 ea/cs
EUR 33
Description: Type: Plate

i-poly Cell Culture Plate (96 well)

IPY-31096 50 ea/cs
EUR 33
Description: Type: Plate

Dimethylsulfoxide (DMSO) for Cell Culture

MBS6120680-100mL 100mL
EUR 395

Dimethylsulfoxide (DMSO) for Cell Culture

MBS6120680-25mL 25mL
EUR 210

Dimethylsulfoxide (DMSO) for Cell Culture

MBS6120680-50mL 50mL
EUR 290

Dimethylsulfoxide (DMSO) for Cell Culture

MBS6120680-5x10mL 5x10mL
EUR 260

Dimethylsulfoxide (DMSO) for Cell Culture

MBS6120680-5x5mL 5x5mL
EUR 210

Plates 96 wells flat bottom for cell culture Cellstar standard Greiner Bio-One - PK100

DD20034 PK100
EUR 256.5

Ultrafiltration Water, cell culture level

PB180518-500mL 500 mL
EUR 15
Description: Supplements & Reagents

Ultrafiltration Water, cell culture level

PB180518 500mL
EUR 15
Description: Others

Ultrafiltration Water, cell culture level

MBS2567648-500mL 500mL
EUR 85

Ultrafiltration Water, cell culture level

MBS2567648-5x500mL 5x500mL
EUR 370

Cell Culture Freezing Medium (NON-FBS)

cAP-22B 50ml
EUR 128.7

3D Cell Culture Gel, medium (Col-Tgel)

P720M-10 10 mL
EUR 389

3D Cell Culture Gel, medium (Col-Tgel)

P720M-2 2 mL
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Gelatin Solution , 0.1%, Cell culture tested

CA087-100 1L
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BD Biocoat 10cm cell culture dish - PK10

354469 PK10
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Mouse Epithelial Cell culture Medium Kit

ABM-TM4285 500 ml Basal medium with the growth factor supplement Ask for price
Description: Epithelial Cell Medium is a complete medium designed for the culture of epithelial cells. It was tested and optimized with epithelial cell growth and proliferation in vitro. It is formulated for use with 5% CO2 and 95% air in a humidified incubator.The medium consists of 500 ml of basal medium (containing essential and non-essential amino acids, vitamins, organic and inorganic compounds, hormones, growth factors, trace minerals), supplemented with epithelial cell growth supplement, antibiotics, and fetal bovine serum.Epithelial Cell Medium Supplement Kit includes:- 0.5 ML RI-Transferrin-Selenium (ITS)- 0.5 ML EGF- 5.0 ML L-Glutamine- 5.0 ML Antibiotic-Antimycotic Solution- 10.0 ML FBS*Before use, add Supplement Kit into 500ml basal medium.

Lipoproteins, Low Density, HP,Cell Culture

12-16-120412-TC-1mg 1mg
EUR 85

Lipoproteins, Low Density, HP,Cell Culture

12-16-120412-TC-5mg 5mg
EUR 350

Human B Cell Culture and Expansion Kit

BC1001-1 1 package
EUR 5200

3D Cell Culture Matrix Duo-Matrix Kit

K519-100 each
EUR 718.8

1M Tris-HCl, pH 9.0 (Cell Culture grade)

IBS-CB077 500 mL
EUR 36.4

i-poly Cell Culture Dish (Ø60 x H15mm)

IPY-10060 500 ea/cs
EUR 49
Description: Type: dish

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IPY-11100 200 ea/cs
EUR 33
Description: Type: dish

Ampicillin sodium salt, cell culture grade

GA0283 5g
EUR 173.88

Ampicillin sodium salt, cell culture grade

GA0283-100 100
EUR 189.8

Ampicillin sodium salt, cell culture grade

GA0283-100G 100 g
EUR 265.2

Ampicillin sodium salt, cell culture grade

GA0283-25 25
EUR 71.1

Ampicillin sodium salt, cell culture grade

GA0283-25G 25 g
EUR 122.4

Ampicillin sodium salt, cell culture grade

GA0283-5 5
EUR 23.8

Ampicillin sodium salt, cell culture grade

GA0283-5G 5 g
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Sodium Bicarbonate. cell culture tested - kg

PM-S2011/1 kg
EUR 10.4

Lipoproteins, High Density, HP, Cell Culture

12-16-080412-TC-1mg 1mg
EUR 80

Lipoproteins, High Density, HP, Cell Culture

12-16-080412-TC-5mg 5mg
EUR 340

CULTURE VESSEL, CULTUREJAR, PT-12

C185 100EA
EUR 48.82

Pluronic® F-127 *Cell culture tested *

20050-10g 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.

40X9mm Cell Culture Plastic Dishes 20/PK

DS001-40 1PK, 20UNIT
EUR 64.18

Cancer Antigen 50 (CA 50) GI cell culture

GWB-3F2336 50000 Unit Ask for price

Sodium Bicarbonate. cell culture tested - 500g

PM-S2011/500 500g
EUR 6.71

60X12mm Cell Culture Plastic Dishes 20/PK

DS001-60 1PK, 20UNIT
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90X15mm Cell Culture Plastic Dishes 10/PK

DS001-90 1PK, 10UNIT
EUR 65.02

Adult Keratinocyte Cell Culture Kit (192627 + 192060)

LO00192627B each
EUR 1015.28

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.