Efficient differentiation between Salmonella Pullorum and Salmonella Gallinarum by a fimH-based PCR-HRM.

IF 2.5 2区农林科学 Q1 VETERINARY SCIENCES Avian Pathology Pub Date : 2025-01-21 DOI:10.1080/03079457.2025.2450840

Di Zhang, Linlin Zhuang, Yi Jiang, Yi Yang, Ming Xu, Xinhong Dou, Jiansen Gong

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Abstract

Salmonella Pullorum (S. Pullorum) and Salmonella Gallinarum (S. Gallinarum) are the biovars of Salmonella enterica serovar Gallinarum that are responsible for pullorum disease and fowl typhoid, respectively, in poultry. Traditional serological methods fail to quickly differentiate between these biovars due to their identical O antigenic factors (O9 and O12). Although single nucleotide polymorphism (SNP)-based methods have been used to distinguish between the biovars, they often lack the required accuracy and effectiveness. In this study, we developed a PCR high resolution melt (PCR-HRM) assay, which targeted a SNP at position 665 of the fimH gene, for rapid differentiation between S. Pullorum and S. Gallinarum. Our method showed 100% specificity and was able to detect as little as 0.033 pg of S. Pullorum DNA and 0.027 pg of S. Gallinarum DNA. The PCR-HRM results for 547 clinical isolates were in complete agreement with traditional serological methods. This PCR-HRM assay significantly reduced identification time and provided high throughput, efficient testing. This makes it a practical and reliable tool for accurate differentiation between S. Pullorum and S. Gallinarum in clinical settings.

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基于荧光定量PCR-HRM的白痢沙门氏菌与鸡痢沙门氏菌的高效鉴别

鸡痢沙门氏菌（S. Pullorum）和鸡痢沙门氏菌（S. Gallinarum）是肠道沙门氏菌血清型鸡痢沙门氏菌的生物变体，分别在家禽中引起鸡痢病和禽伤寒。传统的血清学方法无法快速区分这些生物变体，因为它们具有相同的O抗原因子（O9和O12）。尽管基于单核苷酸多态性（SNP）的方法已被用于区分生物变体，但它们往往缺乏所需的准确性和有效性。在这项研究中，我们开发了一种PCR高分辨率熔体（PCR- hrm）检测方法，该方法针对fimH基因665位的一个SNP，用于快速区分S. Pullorum和S. Gallinarum。我们的方法具有100%的特异性，能够检测到低至0.033 pg的白痢沙门氏菌DNA和0.027 pg的鸡肋沙门氏菌DNA。547株临床分离株的PCR-HRM结果与传统血清学方法完全一致。该PCR-HRM分析显著缩短了鉴定时间，提供了高通量、高效的检测。这使得它成为一个实用和可靠的工具，准确区分鸡鸡沙门氏菌和鸡鸡沙门氏菌在临床设置。

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来源期刊

Avian Pathology 农林科学-兽医学

CiteScore

4.50

自引率

10.70%

发文量

审稿时长

1 months

期刊介绍： Avian Pathology is the official journal of the World Veterinary Poultry Association and, since its first publication in 1972, has been a leading international journal for poultry disease scientists. It publishes material relevant to the entire field of infectious and non-infectious diseases of poultry and other birds. Accepted manuscripts will contribute novel data of interest to an international readership and will add significantly to knowledge and understanding of diseases, old or new. Subject areas include pathology, diagnosis, detection and characterisation of pathogens, infections of possible zoonotic importance, epidemiology, innate and immune responses, vaccines, gene sequences, genetics in relation to disease and physiological and biochemical changes in response to disease. First and subsequent reports of well-recognized diseases within a country are not acceptable unless they also include substantial new information about the disease or pathogen. Manuscripts on wild or pet birds should describe disease or pathogens in a significant number of birds, recognizing/suggesting serious potential impact on that species or that the disease or pathogen is of demonstrable relevance to poultry. Manuscripts on food-borne microorganisms acquired during or after processing, and those that catalogue the occurrence or properties of microorganisms, are unlikely to be considered for publication in the absence of data linking them to avian disease.