Skånseng Beate, Kaldhusdal Magne, Rudi Knut
MATFORSK, Norwegian Food Research Institute, Osloveien 1, N-1430 As, Norway.
Mol Cell Probes. 2006 Oct;20(5):269-79. doi: 10.1016/j.mcp.2006.02.001. Epub 2006 Mar 6.
We compared the colonisation of the chicken gut by the two important pathogens Campylobacter jejuni (frequent food-borne pathogen) and alpha-toxin gene containing Clostridium perfringens (causative agent of necrotic enteritis in chickens) using a new high-throughput automated DNA purification method for microbial biodiversity analyses. The method gave high reproducibility (standard deviation of 1.1 C(T)-values for a universal 16S rDNA real-time PCR), and inhibition was observed in only 0.9% of the individual DNA purifications (n = 753). We analysed 253 randomly collected chicken caecal samples (sampled in 2001 and 2003) from Norwegian chicken flocks by real-time quantitative PCR. Our results showed positive correlation (P = 0.009) in chicken caecal colonisation between C. jejuni and Cl. perfringens. We also found that there was a significant underrepresentation (P = 0.008) of chickens containing high levels of Cl. perfringens and low levels of C. jejuni. This indicates a possible interaction between these bacteria. Potential interaction between pathogens and other bacteria in the gut will certainly be important research fields in the future. As demonstrated here, the development of new tools for high-throughput analyses will be of key importance for these studies.
我们使用一种用于微生物多样性分析的新型高通量自动化DNA纯化方法,比较了空肠弯曲杆菌(常见食源性病原体)和含有α毒素基因的产气荚膜梭菌(鸡坏死性肠炎病原体)在鸡肠道中的定殖情况。该方法具有高重现性(通用16S rDNA实时PCR的C(T)值标准偏差为1.1),并且在仅0.9%的个体DNA纯化中观察到抑制作用(n = 753)。我们通过实时定量PCR分析了从挪威鸡群中随机收集的253份鸡盲肠样本(于2001年和2003年采集)。我们的结果显示,空肠弯曲杆菌和产气荚膜梭菌在鸡盲肠定殖之间呈正相关(P = 0.009)。我们还发现,含有高水平产气荚膜梭菌和低水平空肠弯曲杆菌的鸡存在显著代表性不足的情况(P = 0.008)。这表明这些细菌之间可能存在相互作用。肠道中病原体与其他细菌之间的潜在相互作用肯定将是未来重要的研究领域。如此处所示,开发用于高通量分析的新工具对于这些研究至关重要。