| 摘要: |
| [目的]对奶牛产后子宫内膜炎致病菌进行16S rRNA序列鉴定.[方法]从产后子宫内膜炎患牛子宫分泌物中分离致病菌,通过细菌培养、纯化、分离、革兰氏染色和生化试验进行初步鉴定.选取代表性菌株11株,利用细菌通用引物,通过PCR方法,对其16S rRNA基因的核苷酸序列进行扩增,将扩增产物与pMD19-T载体连接构建克隆载体,经PCR和双酶切鉴定正确后测序,测序结果与GenBank中已注册菌株的16S rRNA基因序列进行比对.[结果]共分离到致病菌株60株,有代表性的11株细菌归类为:SD01为琼氏不动杆菌,SD02为粪肠球菌,SD03为金黄色葡萄球菌,SD04为中间葡萄球菌,SD05为溶血葡萄球菌,SD06为鲁菲不动杆菌,SD07为无乳链球菌,SD08为芽孢杆菌,SD09为枯草芽孢杆菌,SD10为大肠杆菌,SD11为假单胞杆菌.[结论]通过国际公认的16S rRNA序列鉴定技术,准确鉴定出引起奶牛子宫内膜炎的致病菌种类,为临床治疗该病提供了理论依据. |
| 关键词: 奶牛 子宫内膜炎 16S rRNA |
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| 基金项目:山东省农科院重大成果培育基金（2006YCG012） |
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| Identification of the pathogenic bacteria from endometritis cow by 16S rRNA sequence analysis |
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| Abstract: |
| 【Objective】 Pathogenic bacteria of the cow puerperal endometritis were identified by the 16S rRNA sequence analysis. 【Method】 baceria were isdated from the secretion of the cows infected with which have puerperal endometritis.Eleven typical strains were selected after culture,purification,separation,gram staining and identification of biochemistry.The universal primer of bacteria was used to amplify the 16S rRNA sequences by PCR.Then the PCR product was linked to the pMD19-T vector to construct cloning vector.The product was sequenced after PCR amplification and double enzyme digestion.The sequences were compared with the 16S rRNA sequences of the strains that have been reqisterced in the GenBank database.【Result】 We have isolated 60 strains,and 11 strains are typical. we classify it as follows:Acinetobacter junii(SD01),Enterococcus faecalis(SD02),Staphylococcus aureus(SD03),Staphylococcus intermedius(SD04),Staphylococcus haemolyticus(SD05),Acinetobacter lwoffii(SD06),Streptococcus agalactiae(SD07),Bacillus(SD08),Bacillus subtilis(SD09),Escherichia coli(SD10),Pseudomonas sp(SD11).【Conclusion】The pathogenic bacteria that cause cow puerperal endometritis have been determind by the internationally recognized method(16S rRNA sequence analysis method).The results of this experiment will a provide theoretical basis for clinical therapy of cow puerperal endometritis. |
| Key words: cow endometritis 16S rRNA |
