Establishment of droplet digital PCR for quantification of virus particles in recombinant adenovirus-vectored SARS-CoV-2 vaccine / 中华微生物学和免疫学杂志
Chinese Journal of Microbiology and Immunology
;
(12): 881-885, 2023.
Article
in Chinese
| WPRIM
| ID: wpr-1029489
ABSTRACT
Objective:
To construct a droplet digital PCR (ddPCR) for quantification of virus particles in recombinant adenovirus-vectored SARS-CoV-2 vaccine.Methods:
The genome of SARS-CoV-2 BA.1 strain was used as the target gene. A set of primer and probe was designed based on the conserved sequence of spike protein, and the ddPCR for quantification of virus particles in recombinant adenovirus-vectored SARS-CoV-2 vaccine was established. The linearity, accuracy, specificity, repeatability and durability of the method were verified.Results:
The optimal annealing temperature for ddPCR was 63℃. When the number of virus particles was in the range of 5×10 5-2×10 7 VP/ml, the linearity and the recovery rate of the method were good; the specificity of the probe and primer was good; the coefficient of variation (CV) values of the repeatability and the intermediate precision were within 10%; the CV value of the durability was within 15%. When comparing ddPCR with qPCR, the CV values were all within 10%.Conclusions:
The established ddPCR had high sensitivity, good stability and strong specificity, suggesting that it could be used for the quantitative detection of virus particles in recombinant adenovirus-vectored SARS-CoV-2 vaccine.
Full text:
Available
Index:
WPRIM (Western Pacific)
Language:
Chinese
Journal:
Chinese Journal of Microbiology and Immunology
Year:
2023
Type:
Article
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