OPTIMASI METODE INHIBISI AKTIVITAS NEURAMINIDASE DAN PENGEMBANGAN RT-PCR UNTUK PENENTUAN SUBTIPE NEURAMINIDASE PADA VAKSIN INFLUENZA
Vaccination is a main step in controlling influenza disease and reducing number of human deaths due to seasonal influenza that occurs every year and pandemic influenza that might occurs anytime. Quality control of influenza vaccine production through neuraminidase subtype determination as required b...
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| Format: | Theses |
| Language: | Indonesia |
| Online Access: | https://digilib.itb.ac.id/gdl/view/79055 |
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| Institution: | Institut Teknologi Bandung |
| Language: | Indonesia |
| Summary: | Vaccination is a main step in controlling influenza disease and reducing number of human deaths due to seasonal influenza that occurs every year and pandemic influenza that might occurs anytime. Quality control of influenza vaccine production through neuraminidase subtype determination as required by WHO was the focus of this study. The purpose of this research is to establish neuraminidase subtype determination method in an Indonesian vaccine company by optimizing neuraminidase activity inhibition method which was adapted from NVI (The Netherlands) and also to apply alternative genetic method using RT-PCR. The samples were bulk monovalent influenza vaccine A/California/7/2009 (H1N1), A/Victoria/210/2009 (H3N2), B/Brisbane/60/2008 (B), and A/Indonesia /5/2005 (H5N1) subtype. Optimization result in first step of neuraminidase activity inhibition method was a parameter to determine sample dilution level which has significant absorbance decrease in sample dilution and A550 curve. Optimization of the criteria for determining subtypes of neuraminidase N1 and N2 was inhibition titer of a spesific antiserum against a sample should be at least two times greater than other antiserum inhibition titer. For neuraminidase subtype determination through RT-PCR, four pairs of specific primers has been designed for the four subtypes. The experimental results indicate that the primers are specific to three subtypes and generate PCR product 455 bp (N1/Cal), 562 bp (N2/Vic), and 305 bp (NB/Bris) in size. Primer for subtype N1/Ind did not show optimal results possibly due to differences in the nucleotide sequence between the sample and data bank. In conclusion, neuraminidase activity inhibition method was optimum to be used for determining neuraminidase subtypes in the vaccine company, while RT-PCR can be used for alternative method.
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