Objective To perform species identification of Brucella isolated from the whole blood of Marmota himalayana by PCR. Methods A total of 245 whole blood samples were collected from M. himalayana in Qinghai province, China in 2016. Serum was isolated, and the Rose Bengal plate test and the tube agglutination test were used to detect brucellosis antibody. The blood culture method was used to perform bacterial culture of the whole blood samples of M. himalayana with positive antibody. The test results were observed and suspected Brucella strains were isolated and purified. Smear microscopy and the bacteriophage lysis test were performed for the purified suspected Brucella strains, nucleic acid was extracted to prepare templates, and then BCSP31-PCR and AMOS-PCR were used to identify Brucella strains. Results One suspected Brucella strain was isolated with the same morphology and staining characteristics as Gram-negative bacilli. The bacteriophage lysis test showed lysis of BK2 and non-lysis of Tb, and the strain could agglutinate with the single-phase specific serum A and M; therefore, the strain was identified as Brucella melitensis type Ⅲ. BCSP31-PCR agarose gel electrophoresis showed a specific target band at 223 bp, and AMOS-PCR agarose gel electrophoresis showed a specific target band at 731 bp; the strain was proved to be B. melitensis. Conclusion A Brucella strain is isolated for the first time from 245 whole blood samples of M. himalayana and is confirmed to be B. melitensis by PCR in Qinghai province.
ZHANG Ai-ping, XU Li-qing, MA Li, LI Ji-quan, XUE Hong-mei, WANG Jian-ling, YANG Xu-xin, XIE Hui, ZHAO Yuan-bo, YU Shou-hong
. Identification of a Brucella strain isolated from Marmota himalayana in Qinghai province, China[J]. Chinese Journal of Vector Biology and Control, 2021
, 32(2)
: 217
-220
.
DOI: 10.11853/j.issn.1003.8280.2021.02.019
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