猪鼻支原体血清抗体间接ELISA检测方法的建立

doi:10.11843/j.issn.0366-6964.2017.10.016

Abstract

Abstract:

Mycoplasma hyorhinis (M. hyorhinis) is a commensal inhabitant in swine. It can cause polyserositis, pneumonia, arthritis and pericarditis in pigs. Because of the high cross-reactivity among M. hyorhinis and other swine mycoplasmas, there is no specific serological diagnostic method reported. In this study, an indirect ELISA was established by using the variable lipoprotein (Vlp) as the coating antigen. By optimizing the reaction conditions, the optimal concentration of coating antigen was 0.312 5 μg·mL^-1 and the serum dilution was 1∶100, and serum reaction time was 30 min. The optimal dilution at 1∶20 000 for the goat anti-pig HRP-IgG was determined, and the reaction time was 30 min. The optimal substrate chromogenic time was 10 min. The cut-off value of the assay was 0.27. In the cross-reactivity experiment, the Vlp-ELISA showed no cross-reactivity with common swine pathogens or other swine mycoplasmas. The coefficients of variation of the inter-assay and intra-assay experiments were both below 10%. To investigate the specificity and sensitivity of this method, the positive and negative sera samples were detected by the Vlp-ELISA. After that, a total of 247 clinical samples and sera from the experimentally-infected pigs were tested by the Vlp-ELISA, and the results showed the same tendency with that of Tween 20 extracted membrane protein-ELISA method. In conclusion, the indirect ELISA established in this study was specificity and stability, which can be widely used in clinical detection.

CLC Number:

S854.43

WANG Jia, XIONG Qi-yan, HUA Li-zhong, LIU Bei-bei, GAN Yuan, FENG Zhi-xin, LIU Mao-jun, WU Yu-zi, SHAO Guo-qing. Establishment of an Indirect ELISA Detection Method for Mycoplasma hyorhinis[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(10): 1932-1938.

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Establishment of an Indirect ELISA Detection Method for Mycoplasma hyorhinis

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