山羊淋巴结炎病原菌的分离鉴定及多重PCR检测方法的建立

doi:10.11843/j.issn.0366-6964.2017.02.016

Abstract

Abstract:

The purpose of this paper was to isolate and identify the pathogenic bacteria of lymphadenitis in goat of Chongqing area and establish the test method. Sterile separation operation was carried out to isolate pathogenic bacteria from fester of the goat. The phylogenetic tree was founded according to the gene sequence of 16S RNA. And the specific primers were designed to establish PCR test method. The result showed that Corynebacterium pseudotuberculosis, Arcanobacterium pyogenes and Staphylococcus aureus were isolated from the samples, and three pairs of specific oligonucleotide primers were respectively designed according to the gene sequence of Phospholipases D, 16S RNA and nuc for multiple PCR detection. The multiple PCR detection showed that a fragment of 507, 1 378 and 278 bp amplified from the mixture genome .The sensitivity test showed that the multiplex PCR could detect minimum detectable concentration of 10³ cfu•mL^-1.The detection of 80 clinical samples indicated that, the detection rates of mixed infections of Corynebacterium pseudotuberculosis and Staphylococcus aureus, Corynebacterium pseudotuberculosis and Arcanobacterium pyogenes, three kinds of bacterias were 12.5%, 3.75% and 7.5%, respectively. Three pathogenic bacterias of lymphadenitis in goat were isolated successfully and the multiplex PCR detection method was detected in this study, it laid a foundation for the research of the prevention and control and the pathogenesis for lymphadenitis in goat.

CLC Number:

XU Guo-yang, FU Li-zhi, YANG Jin-long, YANG Liu,XU Deng-feng, ZHANG Yi-fan, SHEN Ke-fei, ZHANG Su-hui. Isolation and Identification of Pathogenic Bacteria of Lymphadenitis in Goat and Establishment of Multiplex PCR Detection Method[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(2): 324-330.

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References

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Isolation and Identification of Pathogenic Bacteria of Lymphadenitis in Goat and Establishment of Multiplex PCR Detection Method

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