A nested-PCR for detection and identification of Brucella vaccine A19 strain
10.3760/cma.j.cn231583-20220331-00101
- VernacularTitle:建立检测与鉴定布鲁氏菌A19疫苗株的巢式PCR方法
- Author:
Guozhong TIAN
1
;
Bo LIU
;
Yuanyuan GUO
;
Liqin SU
;
Bike ZHANG
Author Information
1. 中国疾病预防控制中心传染病预防控制所，北京 102206
- Keywords:
Brucella;
Vaccine A19 strain;
Nested-PCR
- From:
Chinese Journal of Endemiology
2023;42(3):196-199
- CountryChina
- Language:Chinese
-
Abstract:
Objective:A nested-PCR assay is developed to detect and identify the genomic DNA of Brucella vaccine A19 strain. Methods:The whole genomic sequences of Brucella vaccine A19 strain and other Brucella spp. strains were compared and analyzed. The primers were designed by nucleotide difference sites. The nested-PCR assay was established to detect and identify Brucella vaccine A19 strain. The genomic DNA of Brucella vaccine A19 strain was extracted and diluted. The diluted template DNA was tested for sensitivity of using nested-PCR assay. And the specificity of nested-PCR assay was tested for the genomic DNA of other Brucella spp. strains and non- Brucella spp. strains. Results:The minimum detection limit of the nested-PCR assay was 3.43 fg. The nested-PCR assay established for amplification of Brucella vaccine A19 strain showed 246 bp electrophoresis bands, while other Brucella spp. strains showed 314 bp electrophoresis bands, and non- Brucella spp. strains did not produce electrophoresis bands. Conclusions:The nested-PCR assay established has the characteristics of high sensitivity and specificity. It can be detected when there is one copy of Brucella vaccine A19 strain genomic DNA in the reaction system. This method is particularly suitable for the detection and identification of trace genomic DNA of Brucella vaccine A19 strain in sample.
