DETERMINATION OF RANGE, QUANTIFICATION LIMIT, AND DETECTION LIMIT IN SARS-COV-2 PSEUDOLENTIVIRUS-BASED NEUTRALIZATIONÂ ASSAY
The efficacy testing of the COVID-19 vaccine can be conducted using a virus neutralization assay. This test requires BSL-3 handling facilities because the wild-type SARS-CoV-2 virus is highly infectious and dangerous. Through pseudotyping, the formed virus cannot replicate, allowing neutralization...
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| Format: | Final Project |
| Language: | Indonesia |
| Online Access: | https://digilib.itb.ac.id/gdl/view/87773 |
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| Institution: | Institut Teknologi Bandung |
| Language: | Indonesia |
| Summary: | The efficacy testing of the COVID-19 vaccine can be conducted using a virus neutralization assay.
This test requires BSL-3 handling facilities because the wild-type SARS-CoV-2 virus is highly infectious
and dangerous. Through pseudotyping, the formed virus cannot replicate, allowing neutralization
tests to be conducted in BSL-2 handling facilities. This study aims to determine the range, limit of
quantification, and limit of detection parameters for the pseudolentivirus-based neutralization assay.
Pseudolentivirus was formed from a lentivirus vector with the surface spike protein of wild-type
SARS-CoV-2. The assay development started with isolating and confirming plasmids for
pseudolentivirus and neutralizing antibodies (NAb) production. Confirmed plasmids for
pseudolentivirus production are transfected into HEK293T cells. The formed pseudolentivirus infects
HEK293T-ACE2 cells and produces luminescence. NAb was also produced by co-transfecting
confirmed plasmids into HEK293T cells, and indirect ELISA results showed an NAb titer of 36.14 ±
1.16 ng/µL. The range of the pseudolentivirus-based SARS-CoV-2 neutralization assay is 450-1800
ng. The neutralization assay has a quantification limit of 450 ng and a detection limit of 257.77 ng.
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