摘要
本研究旨在将ERIC-PCR基因指纹图谱技术应用于兔粪微生物的分析中,建立一种快速分析检测兔肠道菌群变化的方法。通过对PCR反应条件的优化确定适合兔粪微生物的ERIC-PCR反应条件,对比兔粪微生物组和兔粪优势菌的ERIC-PCR DNA指纹图谱差异,从而分析兔粪微生物ERIC指纹图谱的特征。结果建立了适合兔粪样品ERIC-PCR分析的反应体系,确定450bp处条带为双歧杆菌特征条带,1 300和4 000bp处为大肠杆菌特征条带;经发酵验证,图谱中的相应条带亮度变化趋势与计数平板所测结果相一致(相对亮度R与相对菌落数lg cfu的相关系数为0.967 6)。本研究优化建立了一种快速分析检测兔粪中菌群变化的ERIC-PCR指纹图谱技术,该技术可较好反映兔粪中优势菌含量的变化情况,为兔肠道疾病的预防、诊断和治疗提供依据。
This study was designed to apply ERIC-PCR genetic fingerprinting technology to analyze the microbes in rabbit manure,and establish a rapid analysis and detection method of rabbit manure bacterium changes.The ERIC-PCR DNA fingerprint of bacteria in rabbit manure was established by optimizing PCR reaction conditions.The differences between DNA fingerprints of microbiome and dominant strains isolated from rabbit manure was compared,and then the characteristics of the ERIC fingerprint of bacteria in rabbit manure was analyzed.The ERIC-PCR reaction system for bacteria in rabbit manure was established.The characteristic bands for Bifidobacteria were located at 450 bp,the characteristic bands for E.coli were located at 1 300 and 4 000 bp.Anaerobic fermentation experiment proved that the trends of the bands＇ brightness were consistent with the results of the colony count（the correlation coefficient between the relative brightness and lg cfu was 0.967 6）.A rapid analysis and detection method of rabbit manure bacterium changes by using ERIC-PCR genetic fingerprinting technology was established.This method could reflect the changes of beneficial bacteria in rabbit manure,thus providing a basis for prevention,diagnosis and treatment of rabbit intestinal diseases.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2016年第4期716-722,共7页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
新乡市泰弘生物有限公司委托项目(2477)
