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

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abx412775-02mg 0.2 mg
EUR 565.00
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CMRL 1066, CIT CULTURE MEDIA

98-304-CV 500 mL/pk
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Anti-Hu CD173 Culture supernatant

99-594-L001 1.0 ml
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FINAL WASH SOLUTION/CULTURE MEDIUM

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ROCKER CELL CULTURE BAG, 20L

91-200-78 1/pk
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91-200-79 1/pk
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6-well Cell Culture Plates

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6-well Cell Culture Plates

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Human CD34+ Cell Culture Medium

P771 NULL
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Mouse ES Cell Culture Media

M16101 500 mL
EUR 1114.00

Tissue Culture Plate, 12 Well

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

516-A-01mg 0,1 mg
EUR 267.50
  • 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
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Description: anti-CD45 RA B-cells, T-cells, NK-cells

5637 cells

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MB49 cells

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H716 cells

C0009022 One Frozen vial
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Hep3B cells

C0015001 One Frozen vial
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HepG2 cells

C0015002 One Frozen vial
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H1703 cells

C0016008 One Frozen vial
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SW1573 cells

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H2228 cells

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H1581 cells

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H2444 cells

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H187 cells

C0016061 One Frozen vial
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C0016067 One Frozen vial
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A673 cells

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EUR 455.00

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H211 cells

C0016078 One Frozen vial
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H146 cells

C0016080 One Frozen vial
EUR 455.00

H889 cells

C0016081 One Frozen vial
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H510A cells

C0016082 One Frozen vial
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H128 cells

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H1048 cells

C0016084 One Frozen vial
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A2780 cells

C0017002 One Frozen vial
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A2780cis cells

C0017003 One Frozen vial
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E3LZ10.7 cells

C0018005 One Frozen vial
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A10.7 cells

C0018006 One Frozen vial
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Vcap cells

C0019001 One Frozen vial
EUR 455.00

C32 cells

C0020002 One Frozen vial
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N87 cells

C0023002 One Frozen vial
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AGS cells

C0023004 One Frozen vial
EUR 455.00

8505C cells

C0025002 One Frozen vial
EUR 660.00

K1 cells

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R2C cells

C0028001 One Frozen vial
EUR 543.00

Y79 cells

C0029001 One Frozen vial
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BeWo cells

C0030001 One Frozen vial
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T11D7e2 cells

C0033001 One Frozen vial
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C0033003 One Frozen vial
EUR 660.00

RD cells

C0035001 One Frozen vial
EUR 455.00

HASM cells

P0001001 One Frozen vial
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HBSM cells

P0007001 One Frozen vial
EUR 777.00

SCBN cells

P0010001 One Frozen vial
EUR 543.00

STO cells

P0011006 One Frozen vial
EUR 455.00

A9 cells

P0011007 One Frozen vial
EUR 543.00

L929 cells

P0011012 One Frozen vial
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MDCKII cells

P0014001 One Frozen vial
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VERO cells

P0014002 One Frozen vial
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MDCK cells

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BJ cells

P0020005 One Frozen vial
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HUVEC cells

P0027001 One Frozen vial
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C2C12 cells

P0028001 One Frozen vial
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ST cells

P0028002 One Frozen vial
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C127 cells

S0006001 One Frozen vial
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hFOB1.19 cells

T0004005 One Frozen vial
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SVGp12 cells

T0005008 One Frozen vial
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BSC40 cells

T0011003 One Frozen vial
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Mv1Lu cells

T0016003 One Frozen vial
EUR 455.00

IMCC3 cells

T0017001 One Frozen vial
EUR 719.00

HPDEC cells

T0018001 One Frozen vial
EUR 777.00

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.