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OBJECTIVE To develop and verify two methods for the detection of size distribution of residual Vero cell DNA in the bulk of vaccine. METHODS Based on capillary electrophoresis and microfluidic chip-based electrophoresis, a method for separation and detection of residual Vero cell DNA fragments in different size which have been purified and fluorescent-labelled was established. The developed method was verified for specificity, accuracy, repeatability, sensitivity, and used for detection of size distribution of residual Vero cell DNA fragments in the bulk of two different vaccines. RESULTS Thirteen DNA fragments of different size were separated at baseline level. The relative deviation in retention time between the control product with 200, 500, 1 000 bp DNA and the reference product were lower than 0.5%. The average relative deviations of retention time by 6 times detection of 13 DNA fragments were 0.72% and 0.11%, respectively. The LODs were 1.5 and 0.1 pg·μL-1, respectively. The blank control showed no influence on the test. The result of the sample added with standard DNA fragments was in accordance with the theoretical expectation. No peak was detected in one kind of vaccine sample, and the other one showed two groups of peaks representing different size. CONCLUSION The developed method has good specificity and high sensitivity. It is convenient and rapid, which is suitable for the determination of size distribution of residual DNA fragments in the bulk of vaccine.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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