Townsend K M, Dawkins H J, Papadimitriou J M
UWA Department of Pathology, Q.E. II Medical Centre, Nedlands, Australia.
Res Vet Sci. 1997 Sep-Oct;63(2):151-5. doi: 10.1016/s0034-5288(97)90009-6.
Amplification of multiple P multocida genomic DNA fragments by outwardly-directed primers based on the repetitive extragenic palindromic (REP) consensus sequence, generated complex profiles in a PCR-based fingerprinting method known as REP-PCR. Polymorphisms within REP-PCR profiles were used to characterise 38 isolates of P multocida. The high degree of homogeneity observed among haemorrhagic septicaemia (HS) strains of serotype B and E provided evidence of a disease-associated REP profile that may serve as a novel method for the identification of HS strains regardless of serotype. REP-PCR profiles of other P multocida serotypes were highly variable, illustrating the potential of this technique for the molecular fingerprinting of fowl cholera or atrophic rhinitis isolates. A specific amplified REP fragment was isolated and used to probe membrane-bound digested P multocida genomic DNA. Hybridisation patterns not only distinguished HS-causing isolates from non-HS P multocida, but also demonstrated a degree of relatedness between HS and HS-like strains.
基于重复外显子回文序列（REP）共有序列，通过向外引物扩增多个多杀性巴氏杆菌基因组DNA片段，在一种称为REP-PCR的基于PCR的指纹识别方法中产生了复杂的图谱。REP-PCR图谱中的多态性被用于鉴定38株多杀性巴氏杆菌。在B型和E型出血性败血症（HS）菌株中观察到的高度同质性提供了一种与疾病相关的REP图谱的证据，这可能成为一种识别HS菌株的新方法，而无需考虑血清型。其他多杀性巴氏杆菌血清型的REP-PCR图谱高度可变，说明了该技术在禽霍乱或萎缩性鼻炎分离株分子指纹识别方面的潜力。分离出一个特定的扩增REP片段，并用于探测膜结合的经消化的多杀性巴氏杆菌基因组DNA。杂交模式不仅区分了引起HS的分离株与非HS多杀性巴氏杆菌，还显示了HS菌株与HS样菌株之间的一定程度的相关性。
