Objective To investigate the application of SYBR GreenⅠ fluorescence quantitative PCR in the rapid detection of Brucella, and to provide a new method for the early detection of brucellosis. Methods Bacterial isolation and culture were performed for two whole blood samples collected from patients with acute brucellosis, 8 whole blood samples collected from patients with chronic brucellosis, and 12 whole blood samples collected from sheep, and suspected Brucella strains were screened by smear microscopy and bacteriophage lysis test. Nucleic acid was extracted, templates were prepared, and then SYBR GreenⅠ fluorescence quantitative PCR was used for the detection of Brucella. Results A total of four suspected Brucella strains were isolated, with the same morphology and staining characteristics as Gram-negative bacilli. The bacteriophage lysis test for strains 1, 2, and 3 showed that BK2 lysed and Tb was not and these three strains were determined as Brucella melitensis; the bacteriophage lysis test for strain 4 yielded negative results. All strains except the strain 4 and EV76 showed a specific absorption peak at 88℃ in the melting curve of fluorescent quantitative PCR, and thus they were confirmed as Brucella. Conclusion SYBR GreenⅠ fluorescence quantitative PCR has the advantages of short time, strong specificity, and good repeatability in the detection of Brucella, and therefore, it holds promise for application in the screening for brucellosis and the rapid detection of Brucella.
ZHANG Ai-ping, MA Li, XIE Hui, YANG Xu-xin, XUE Hong-mei, ZHAO Zhi-jun, LI Ji-quan, YU Shou-hong, ZHAO Zhong-zhi, XU Li-qing
. Application of SYBR GreenⅠfluorescence quantitative PCR in the rapid detection of Brucella[J]. Chinese Journal of Vector Biology and Control, 2020
, 31(6)
: 695
-698
.
DOI: 10.11853/j.issn.1003.8280.2020.06.014
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