Gong Jiansen, Iduu Nneka Vivian, Zhang Di, Chenoweth Kelly, Wei Lanjing, Yang Yi, Dou Xinhong, Wang Chengming
Key Laboratory for Poultry Genetics and Breeding of Jiangsu Province, Jiangsu Institute of Poultry Sciences, Yangzhou 225125, China.
Department of Pathobiology, College of Veterinary Medicine, Auburn University, Auburn, AL 36849, USA.
Microorganisms. 2024 Sep 2;12(9):1815. doi: 10.3390/microorganisms12091815.
Pullorum ( Pullorum) and Gallinarum ( Gallinarum) are two biovars of serovar Gallinarum, responsible for pullorum disease and fowl typhoid, which are the most prevalent and pathogenic forms of salmonellosis in poultry in developing countries. Traditional differentiation methods for . Pullorum and . Gallinarum are based on distinct clinical manifestations and biochemical traits, given their indistinguishable nature via serological assays alone. Molecular differentiation methods such as allele-specific PCR and dual PCR combined with gel electrophoresis or enzyme digestion have also been used to discriminate . Pullorum and . Gallinarum, but the detection efficiency is not high. This investigation introduces a Fluorescence Resonance Energy Transfer (FRET) PCR assay targeting the gene, exclusively found in specific serovars such as . Pullorum and . Gallinarum, and exhibiting conserved single-nucleotide polymorphisms across these two biovars. High-resolution melting curve analysis demonstrates distinct dissolution profiles, facilitating the precise discrimination of . Pullorum and . Gallinarum. This FRET-PCR assay exhibits a detection limit of 10 copies per reaction and has been rigorously validated utilizing 17 reference strains and 39 clinical isolates. The innovation presented herein provides a valuable tool for the rapid differentiation of . Pullorum and . Gallinarum, thereby enhancing diagnostic efficiency and molecular surveillance of poultry . The developed -targeting FRET-PCR assay presents a promising alternative to current cumbersome and time-consuming diagnostic modalities, offering significant potential for expedited identification and control of in poultry and mitigating economic losses associated with contamination in poultry production.
鸡白痢（Pullorum）和鸡伤寒（Gallinarum）是鸡沙门氏菌血清型的两个生物变种，分别引起鸡白痢和禽伤寒，这是发展中国家家禽沙门氏菌病最普遍且致病性最强的形式。鉴于仅通过血清学检测无法区分鸡白痢和鸡伤寒，传统的区分方法基于不同的临床表现和生化特性。分子区分方法如等位基因特异性PCR以及结合凝胶电泳或酶切的双重PCR也已用于鉴别鸡白痢和鸡伤寒，但检测效率不高。本研究引入了一种针对该基因的荧光共振能量转移（FRET）PCR检测方法，该基因仅在鸡白痢和鸡伤寒等特定血清型中发现，并且在这两个生物变种中呈现保守的单核苷酸多态性。高分辨率熔解曲线分析显示出明显的解链曲线，有助于精确鉴别鸡白痢和鸡伤寒。这种FRET-PCR检测方法的检测限为每个反应10个拷贝，并已利用17株参考菌株和39株临床分离株进行了严格验证。本文提出的创新为快速区分鸡白痢和鸡伤寒提供了一种有价值的工具，从而提高了家禽诊断效率和分子监测水平。所开发的针对该基因的FRET-PCR检测方法是当前繁琐且耗时的诊断方式的一种有前景的替代方法，在加快家禽中该病菌的鉴定和控制以及减轻家禽生产中该病菌污染造成的经济损失方面具有巨大潜力。
