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AIMS OF THE STUDY
Virucidal assay of a test article
against NIBRG-14 [H5N1] Influenza and Urbani SARS virus at four
concentrations for four incubation time points.
TEST
ARTICLE
Greenbridge Clinical Biocide Cleaner
supplied at double strength
DILUTIONS
AND TIME POINTS
The test article was tested at the
following final dilutions of the single strength formulation;
For SARS; neat (single strength), 1/5, 1/50, 1/500 For the NIBRG-14
assay; neat (single strength), 1/10, 1/100 1/1000 The time points
measured were; 0.5, 2, 5 and 10 minutes.
VIRUS
TITRES AND DILUTIONS USED IN ASSAYS
Influenza NIBRG-14:
stock titre = 6 – 8.5 TCID50/ml used at dilution
Urbani SARS virus: stock titre = 5.5-6.0 TCID50/ml
used neat
PROCEDURES
USED INCLUDING SOP NUMBERS:
1. The determination
of TCID50 using the Karber calculation (SOP VA023-02);
2. Viable cell count using the trypan blue exclusion
assay VA029-02);
3. Titration of samples containing Influenza
virus on MDCK (SOP VA016-01);
4. Haemagglutination Assay (SOP VA018-02)
5. Titration of SARS virus on C1008 cells (virus
is titrated fold down a 96-well plate of C1008 cells and the plate
incubated at 37°C and 5% CO2 for up to 5 days).
SUMMARY
NIBRG-14 [H5N1] Influenza
The test article was found to be virucidal against NIBIRG-14 [H5N1].
at all the dilutions and timepoints tested. Significant reduction
of viral titre (>1 –log10) reduction in virus was observed
at the neat and the 1/10, 1/100, and 1/1000 diluted test article
samples. At the 1/1000 dilution the test article was found to
significantly reduce virus only at the 0.5 minute time point.
The 10, 5 and 2 minute time points were observed to reduce virus,
this reduction was not significant (<1 log10).
SARS Urbani
Virus
Toxicity was observed against the C1008 cells with the neat test
article. This toxicity prevented assessment of virucidal activity
at this concentration. The test compound was found to be virucidal
against SARS Urbani at the 1/5, 1/50 and 1/500 dilutions.
Significant reduction (>1 –log10) reduction in virus
was observed at all the 1/5 and 1/50 test dilutions. The test
article was found to be virucidal at the 1/500 dilution at the
5, 2 and 0.5 minute timepoints. Virucidal activity was not observed
at the 10 minute incubation timepoint at the 1/500 dilution.
VIRUCIDAL ASSAY WITH INFLUENZA NIBRG-14 [H5N1]
*limit of assay due to toxicity
VIRUCIDAL ASSAY WITH URBANI SARS VIRUS
*limit of assay due to toxicity
CONCLUSION
Toxicity was evident at the single
strength concentration on both the MDCK and C1008 cells at the
0.5, 2, 5 and 10 minute time points. Toxicity was also evident
at the 1/5 and 1/50 dilutions on the C1088 cells at the 0.5, 2,
5 and 10 minute time points.
The presence of toxicity masks any observations of viral CPE where
it is observed and also
affects viral growth.
Influenza NIBRG-14 [H5N1]
The highest reduction in viral titre observed was a ≥5.7
reduction (>99.9%) of the Influenza NIBRG-14 [H5N1] virus at
the 1/10 dilution of the single strength formulation at the 0.5,
2, 5 and 10 minute time points.
Urbani SARS
The highest reduction in viral titre observed was a ≥2.5
(>99.5%) reduction of Urbani SARS virus at the 1/50 dilution
of the single strength formulation for 0.5, 2, 5 and 10 minute
time points.
ARCHIVE
STATEMENT
The dedicated laboratory books, study
protocol and final reports, together with any other relevant information,
will be held at the Retroscreen Virology Ltd. premises, Medical
Sciences Building, 327 Mile End Road, London, E1 4NS secure archive
under the
supervision of the Archivist for 12 months from the issuing of
the final report at no charge. After 12 months, storage may continue
at the cost of £50.00 plus VAT per annum per box (W x D
x H; 418 MM x 710 MM x 280 MM) payable in advance or at the discretion
of the customer the material may be returned to the customer.
SAMPLE STORAGE
All samples provided by the sponsor
and all samples generated during the Research Project will be
disposed of three months after completion of the Research Project
and the issue of the final report unless otherwise requested by
the Customer. Storage costs are £10.00 plus VAT per box
(81 9 x 9 samples) per month. Retroscreen Virology Ltd. reserves
the exclusive rights on a small proportion of any samples recovered.
STATEMENT OF COMPLIANCE WITH GOOD LABORATORY PRACTICE REGULATIONS
The study described in this report was conducted in compliance
with the following Good Laboratory Practice standards and I consider
the data to be valid.
• The United Kingdom Good Laboratory Practice Regulations
1999 Statutory Instrument No. 3106.
• OECD Principles of Good Laboratory Practice, (Revised
1997).
Miss Shobana Balasingam
QUALITY ASSURANCE STATEMENT
Quality Assurance has audited this report. The methods, practices
and procedures reported herein are an accurate description of
those employed at Retroscreen Virology Ltd during the course of
the study. Observations and results presented in this final report
form a true and accurate representation of the raw data generated
during the conduct of the study at Retroscreen Virology Ltd.
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