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To conduct genetic typing of Mycobacterium tuberculosis (MTB) in Guizhou Province using the MLVA technique based on 24 VNTR loci, to identify VNTR typing loci suitable for Guizhou MTB strains, and to establish a high-resolution, user-friendly MLVA typing model for MTB. This model will provide technical means for tracing the sources, transmission, and prevention and control of tuberculosis in Guizhou Province.
A total of 124 MTB strains were collected from Guizhou Province. PCR amplification was performed using 24 internationally recognized VNTR loci for MTB, followed by analysis of the amplification products using capillary electrophoresis. The number of repeats at each locus was counted, and allele polymorphism and the discriminatory index for each locus were calculated. Clustering analysis of the tested strains was conducted using BIONUMERICS 8.0 to explore the typing characteristics of MTB in Guizhou Province based on 24-VNTR, 15-VNTR, 12-VNTR, and 8-VNTR.
The allele polymorphism and discriminatory index of the 24 VNTR loci indicated that seven loci, including QUB-11b, had high discriminatory power, eleven loci, including MIRU-10, had moderate power, and six loci, including MIRU-20, had low power. Furthermore, clustering analysis revealed that the 24-VNTR typing exhibited the highest discriminatory ability at 95.16% (118/124), approaching the level of single strain identification, followed by 15-VNTR at 91.13% (113/124), 12-VNTR at 90.32% (112/124), and 8-VNTR at 83.06% (103/124). All four VNTR typing models successfully categorized the 124 MTB strains into four clusters.
This study identified six high-resolution loci, including QUB-11b, as the preferred VNTR loci for MLVA typing of MTB strains in Guizhou Province, with other loci ranked according to their discriminatory power as supplementary typing methods. The 24-VNTR, 15-VNTR, and 12-VNTR methods demonstrated high typing capabilities, providing technical support for molecular tracing of tuberculosis in Guizhou Province.
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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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