Identification by 16S rDNA fragment amplification and determination of genetic diversity by random amplified polymorphic DNA analysis of Pasteurella pneumotropica isolated from laboratory rodents.
Identification by 16S rDNA fragment amplification and determination of genetic diversity by random amplified polymorphic DNA analysis of Pasteurella pneumotropica isolated from laboratory rodents.
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通过 16S rDNA 片段扩增进行鉴定，并通过随机扩增多态性 DNA 分析测定从实验室啮齿动物中分离的嗜肺巴氏杆菌。
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发表时间：
1999
期刊：
影响因子：
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通讯作者：
Y. Richard
中科院分区：
文献类型：
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作者：
A. Kodjo;L. Villard;F. Veillet;F. Escande;E. Borges;F. Maurin;J. Bonnod;Y. Richard
嗜肺巴氏杆菌是一种机会性细菌，经常从各种实验室啮齿动物的菌落中分离出来。该物种的鉴定，包括将其分化为两种不同的生物型（Jawetz 和 Heyl），通常基于使用传统的细菌学方法。在本研究中，开发了 16S rDNA 片段扩增程序，作为鉴定和区分肺炎假单胞菌的替代方法。聚合酶链式反应 (PCR) 产物是两个独特的片段，分别为 Jawetz 和 Heyl 生物型特异的 937 bp 和 564 bp。 PCR 产物的特异性可以通过 EcoRI 切割来实现，从而为每个扩增产物产生 596 + 341-bp 和 346 + 218-bp 片段。使用该程序确认了 34 种现场分离株的鉴定，并能够明确鉴定一些细菌学检查无法完成的菌株。与生物型 Heyl 菌株相比，接受随机扩增多态性 DNA (RAPD) 分析的现场分离株在生物型 Jawetz 菌株中具有较高的遗传多样性。总之，RAPD 可以代表鉴定 Heyl 生物型模糊菌株的另一种方法，也是鉴定嗜肺假单胞菌生物型 Jawetz 菌株的有价值的流行病学工具。
Pasteurella pneumotropica is an opportunistic bacterium frequently isolated from colonies of various laboratory rodents. Identification of this species, including its differentiation into two distinct biotypes (Jawetz and Heyl), is usually based on the use of conventional bacteriologic methods. In this study, a 16S rDNA fragment amplification procedure was developed for use as an alternative method for identification and differentiation of P. pneumotropica. Polymerase chain reaction (PCR) products were two distinctive fragments of 937 and 564 bp specific for biotypes Jawetz and Heyl, respectively. Specificity of PCR products could be achieved by EcoRI cleavage, leading to 596 plus 341-bp and 346 plus 218-bp fragments for each of the amplification products. Use of this procedure confirmed identification of 34 field isolates and allowed definitive identification of some strains that could not have been done by use of bacteriologic examinations. Field isolates subjected to random amplified polymorphic DNA (RAPD) analysis had high genetic diversity among biotype Jawetz strains in contrast to biotype Heyl strains. In conclusion, RAPD could represent an additional means for identification of ambiguous strains of biotype Heyl and a valuable epidemiologic tool for identification of biotype Jawetz strains of P. pneumotropica.