牦牛肠道病毒间接免疫荧光检测方法的建立与初步应用

doi:10.11843/j.issn.0366-6964.2016.12.015

Abstract

Abstract:

To establish an method for rapid and accurate detection of the yak enterovirus, the yak enterovirus SWUN-AB001 obtained by our laboratory was used to immunize healthy rabbits, and then establish an indirect immunofluorescent assay (IFA) by using the rabbit hyperimmune serum. And the working conditions and time were screened and optimized. The best dilution and working time of hyperimmune serum in the IFA method were 1:100 and 30 minutes, and these of FITC conjugated goat anti-rabbit IgG were 1:800 and 45 minutes. Specificity of bright green fluorescent particles about the yak enterovirus were clearly visible; it can′t react with Bovine viral diarrhea virus, Rotavirus, E. coli, Salmonella, Streptococcus, Staphylococcus and Pasteurella; the minimum concentration of SWUN-AB001 can be detected by the method was about 20TCID₅₀•mL^-1. Forty clinical diarrhea samples were detected by using the RT-PCR and the IFA, and there was no difference in the experiment results between the RT-PCR and the IFA (P=0.329). The results indicate that the method has strong specificity, good repeatability and high sensitivity, it can be used in study of epidemiological investigation, clinical diagnosis and laid a foundation of the fast diagnostic reagents.

CLC Number:

S852.659.6

HE Huan, YE Yong-gang, CHEN Xin-nuo, WANG Bin-xing, QIN Si-nan, LIU Yan, TANG Cheng, ZHANG Bin. Development and Preliminary Application of an Indirect Immunofluorescent Assay of the Yak Enterovirus[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(12): 2450-2456.

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Development and Preliminary Application of an Indirect Immunofluorescent Assay of the Yak Enterovirus

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