Department of Biotechnology and Genetic Engineering, Philadelphia University, Amman, Jordan.
Braz J Microbiol. 2024 Sep;55(3):2547-2556. doi: 10.1007/s42770-024-01437-4. Epub 2024 Jul 9.
Campylobacter is gram-negative bacteria considered the predominant genera isolated from poultry samples and associated with gastroenteritis. Due to the problems in conventional cultural methods of time-consuming and technically demanding requirements, a rapid and feasible method for their identification and discrimination of the closely related spp. Including Campylobacter coli, Campylobacter fetus, and Campylobacter jejuni is needed. This study analyzes the chicken and sheep meats samples (n = 125) using culture and pre-enrichment-based Quadraplex real-time PCR by targeting OrfA, CstA, HipO, and 16 S rRNA genes of C. coli, C. fetus, C. jejuni and Campylobacter spp. Respectively. The analysis of 125 chicken and sheep meat samples by culture and real-time PCR showed high concordance between the results of the two methods. The present study show high prevalence of Campylobacter species (35% and 32% from chicken and meat respectively) of which C. jejuni were the most abundant. Reaction efficiencies were between 90 and 110%, and detect as low as 8.9 fg in C. jejuni. The need for quick detection and discrimination methods in sheep and chicken meat can be met using the described Quadraplex real-time PCR methodology.
空肠弯曲菌是一种革兰氏阴性菌，被认为是从家禽样本中分离出来的主要属，与肠胃炎有关。由于传统的文化方法存在耗时和技术要求高的问题，因此需要一种快速且可行的方法来鉴定和区分密切相关的 spp. ，包括大肠弯曲菌、胎儿弯曲菌和空肠弯曲菌。本研究使用基于培养和预富集的四重实时 PCR 方法，针对 C. coli、C. fetus、C. jejuni 和 Campylobacter spp. 的 OrfA、CstA、HipO 和 16S rRNA 基因，对 125 份鸡肉和羊肉样本（n=125）进行了分析。对 125 份鸡肉和羊肉样本的培养和实时 PCR 分析表明，两种方法的结果高度一致。本研究表明，弯曲菌属的流行率很高（鸡肉和羊肉分别为 35%和 32%），其中空肠弯曲菌最为丰富。反应效率在 90%至 110%之间，在空肠弯曲菌中可检测到低至 8.9fg 的水平。该描述的四重实时 PCR 方法可以满足快速检测和区分羊肉和鸡肉中弯曲菌的需要。
