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Different library construction methods, combined with high-throughput sequencing methods, were used to understand the genetic characteristics of the virus of the first confirmed imported monkeypox case in mainland China, and to compare the advantages and disadvantages of different library types.
The herpetic fluids and nasopharyngeal swabs of the first imported monkeypox case in the Chinese mainland were used as samples. After nucleic acid extraction and quantification, the library was directly constructed or the amplicon library was constructed using the monkeypox virus whole genome capture kit, and the whole genome sequence of the virus was obtained by high-throughput sequencing. Combined with 34 monkeypox virus sequences downloaded from the NCBI and GISAID databases, a phylogenetic evolutionary tree was constructed using vaccinia, variola, and cowpox sequences as out groups.
The whole genome sequence of the virus was obtained by both library construction methods, named hMpxV/China/CQ-CQCDC-001/2022, which belongs to the monkeypox virus branch IIb, located in the same branch as hMpxV/Germany/BE-ChVir28656/2022 belonging to the IIb B.1 branch. Metagenomic libraries have more uniform sequence coverage than amplicon libraries, but the sequencing depth is lower and the effective data volume accounts for less.
Clade IIb B.1 monkeypox viruses have already been introduced into the mainland China, and the sequencing of subsequently discovered monkeypox viruses should be based on the sample size and sequencing timeframe, sequencing cost-effectiveness ratio, and other factors to choose the appropriate library construction method.
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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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