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.

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

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ExoStep Cell Culture

ExoS-25-C9 25 test
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ExoStepTM Culture + Standard

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PetakaG3 FLAT Culture Devices

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abx412775-02mg 0.2 mg
EUR 565
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Human MEP Cell Culture Medium (human megakaryocyte–erythroid progenitor cell culture medium)

P770 NULL
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SP31171 1CS, 500UNIT
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6-well Cell Culture Plates

abx090688-50plates 50 plates
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6-well Cell Culture Plates

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ROCKER CELL CULTURE BAG, 20L

91-200-78 1/pk
EUR 411
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91-200-79 1/pk
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91-200-80 1/pk
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CMRL 1066, CIT CULTURE MEDIA

98-304-CV 500 mL/pk
EUR 65
Description: Specialty Media; Islet products

NEONATAL PORCINE ISLET CULTURE MEDIUM

98-343-CV 500 mL/pk
EUR 111
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Anti-Hu CD173 Culture supernatant

99-594-L001 1.0 ml
EUR 122

FINAL WASH SOLUTION/CULTURE MEDIUM

99-785-CV 500 mL/pk
EUR 164
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Mouse ES Cell Culture Media

M16101 500 mL
EUR 1114

Human CD34+ Cell Culture Medium

P771 NULL
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UltraPure Water, Cell Culture Tested

W9200-045 450ml
EUR 80

Tissue Culture Plate, 12 Well

TCP20-12 1 UNIT
EUR 53.48
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anti-CD45 RA B-cells, T-cells, NK-cells

516-A-01mg 0,1 mg
EUR 267.5
  • Category: Antibody, Signal Transduction Antibodies, mAb
Description: anti-CD45 RA B-cells, T-cells, NK-cells

anti-CD45 RA B-cells, T-cells, NK-cells

516-A-1000ug 1000 ug
EUR 1282.5
  • Category: Antibody, Signal Transduction Antibodies, mAb
Description: anti-CD45 RA B-cells, T-cells, NK-cells

5637 cells

C0002001 One Frozen vial
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MB49 cells

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C0009022 One Frozen vial
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C0013003 One Frozen vial
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Hep3B cells

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HepG2 cells

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H460 cells

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A427 cells

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H23 cells

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SW1573 cells

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H520 cells

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H226 cells

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H1975 cells

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H2228 cells

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EUR 543

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EUR 543

H526 cells

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EUR 455

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C0016067 One Frozen vial
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C0016068 One Frozen vial
EUR 695

SW1271 cells

C0016073 One Frozen vial
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A673 cells

C0016074 One Frozen vial
EUR 455

H524 cells

C0016077 One Frozen vial
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H211 cells

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H146 cells

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H889 cells

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H1048 cells

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A2780cis cells

C0017003 One Frozen vial
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E3LZ10.7 cells

C0018005 One Frozen vial
EUR 660

A10.7 cells

C0018006 One Frozen vial
EUR 660

Vcap cells

C0019001 One Frozen vial
EUR 455

C32 cells

C0020002 One Frozen vial
EUR 455

N87 cells

C0023002 One Frozen vial
EUR 455

AGS cells

C0023004 One Frozen vial
EUR 455

8505C cells

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EUR 660

K1 cells

C0025003 One Frozen vial
EUR 660

R2C cells

C0028001 One Frozen vial
EUR 543

Y79 cells

C0029001 One Frozen vial
EUR 455

BeWo cells

C0030001 One Frozen vial
EUR 455

T11D7e2 cells

C0033001 One Frozen vial
EUR 602

NB3 cells

C0033003 One Frozen vial
EUR 660

RD cells

C0035001 One Frozen vial
EUR 455

HASM cells

P0001001 One Frozen vial
EUR 777

HBSM cells

P0007001 One Frozen vial
EUR 777

SCBN cells

P0010001 One Frozen vial
EUR 543

STO cells

P0011006 One Frozen vial
EUR 455

A9 cells

P0011007 One Frozen vial
EUR 543

L929 cells

P0011012 One Frozen vial
EUR 455

MDCKII cells

P0014001 One Frozen vial
EUR 455

VERO cells

P0014002 One Frozen vial
EUR 455

MDCK cells

P0014003 One Frozen vial
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BJ cells

P0020005 One Frozen vial
EUR 455

HUVEC cells

P0027001 One Frozen vial
EUR 485

C2C12 cells

P0028001 One Frozen vial
EUR 455

ST cells

P0028002 One Frozen vial
EUR 543

C127 cells

S0006001 One Frozen vial
EUR 485

hFOB1.19 cells

T0004005 One Frozen vial
EUR 455

SVGp12 cells

T0005008 One Frozen vial
EUR 543

BSC40 cells

T0011003 One Frozen vial
EUR 543

Mv1Lu cells

T0016003 One Frozen vial
EUR 455

IMCC3 cells

T0017001 One Frozen vial
EUR 719

HPDEC cells

T0018001 One Frozen vial
EUR 777

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.