绵羊痒螨兔亚种MMP2基因的转录水平及其重组蛋白间接ELISA的建立

doi:10.11843/j.issn.0366-6964.2022.02.023

Abstract

Abstract: The object of this study was to analyze the transcription profiles of the MMP2 in Psoroptes ovis var. cuniculi (PocMMP2), and to establish an indirect ELISA method (iELISA) for detecting IgG antibody against recombinant PocMMP2 (rPocMMP2). The transcription profiles of PocMMP2 across four life-cycle stages, "fed" and "starved" mites were investigated by quantitative real-time PCR. The rPocMMP2 was obtained in Escherichia coli expression system, and this recombinant protein was used to develop rPocMMP2-iELISA by the checkerboard titration tests. Finally, the field investigation using this iELISA was performed for analysis of 86 sera samples collected from a rabbit farm with a previous history of psoroptic mange in Chengdu. Results were as follows:PocMMP2 was expressed across four life-cycle stages, and expression level of MMP2 was significantly higher in "starved" mites than in "fed" mites (P<0.05). The rPocMMP2 was mainly presented in inclusion bodies with the size of~72 ku. The cut-off value for rPocMMP2-iELISA was 0.223, the sensitivity and specificity for the test were 86.0% and 83.3%,respectively. Additionally, the area under the receiver operating curve (AUC) for the test was 0.876,and both the intra-and inter-coefficients of variation were less than 10.00%. Further field study showed 74.42% (64/86) seropositive rate, which was higher than etiological detection rate (10.47%, 9/86). In summary, PocMMP2 might play an important role in the developmental physiology and the invasion process. The rPocMMP2-iELISA appeared good sensitivity, specificity and high repeatability, and could be used to detect serum antibody of psoroptic mange in rabbits.

Key words: Psoroptes ovis var. cuniculi, MMP2, transcription level, prokaryotic expression, indirect ELISA

CLC Number:

S852.746

GE You, ZHENG Jing, PU Shanqiu, ZHOU Yu, HUANG Cuirui, YANG Guangyou, XIE Yue, HE Ran, XU Jing, GU Xiaobin. The Transcription Level of MMP2 in Psoroptes ovis var. cuniculi and Establishment of the Indirect ELISA Based on Its Recombinant Protein[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(2): 567-575.

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The Transcription Level of MMP2 in Psoroptes ovis var. cuniculi and Establishment of the Indirect ELISA Based on Its Recombinant Protein

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