Multiple PCR assay based on the cigR gene for detection of Salmonella spp. and Salmonella Pullorum/Gallinarum identification.

Salmonella spp. are important zoonotic pathogens that are responsible for severe diseases in both animals and humans. Salmonella enterica subsp. enterica serovar Gallinarum biovar Gallinarum (S. Gallinarum) and biovar Pullorum (S. Pullorum) are typical infectious pathogens detected in the chicken industry that have caused great economic losses. To facilitate their detection and prevent contamination, we developed a rapid multiple PCR method, which can simultaneously detect Salmonella spp. and further identify the biovars S. Pullorum/Gallinarum. This PCR detection method is based on the cigR gene, which is conserved among Salmonella spp. but has a 42-bp deletion in S. Pullorum/Gallinarum. The specificity and sensitivity of the PCR assay was evaluated with 41 different strains: 34 Salmonella strains, including 5 S. Pullorum/Gallinarum strains, and 7 non-Salmonella strains. The lower limit of detection was 8.15 pg of S. Pullorum (S06004) genomic DNA and 20 cfu in PCR, which shows a great sensitivity. In addition, this method was applied to detect or identify Salmonella from processing chicken liver and egg samples, and the results corresponded to those obtained from serotype analysis using the conventional slide agglutination test. Overall, the new cigR-based PCR assay is efficient and practical for Salmonella detection and S. Pullorum/Gallinarum identification and will greatly reduce the workload of epidemiologic investigation.

Prevalence, characteristics, and antimicrobial resistance patterns of Salmonella in retail pork in Jiangsu province, eastern China.

Salmonella is commonly isolated from raw pork and is a leading cause of foodborne illness. Because China has the highest rate of pork consumption and the largest number of pig breeding facilities in the world, an epidemiological analysis of Salmonella species from pork in China is warranted. In this study, pork samples (n = 1,096) were collected from 20 major free markets in four cities of Jiangsu province from August 2010 to December 2012. A total of 163 Salmonella isolates were recovered from 154 Salmonella-positive samples. Among 14 Salmonella serovars identified, Derby (47.9%) was most prevalent, followed by Typhimurium (10.4%), Meleagridis (9.2%), Anatum (8.6%), and London (6.7%). Antimicrobial sensitivity testing revealed that 134 (82.2%) of the isolates were resistant to at least one antimicrobial agent, and 41 (25.2%) were resistant to more than three antimicrobials. The highest resistance was to tetracycline (66.3% of isolates) followed by ampicillin (39.9%), trimethoprim-sulfamethoxazole (31.3%), and nalidixic acid (30.1%). Multilocus sequence typing analysis revealed 14 sequence type (ST) patterns; ST40 was the most common (77 isolates) followed by ST64 (19 isolates). Our research revealed a high prevalence of Salmonella in retail pork. Diversity among the Salmonella isolates was high in terms of serovar and genotype, and multidrug resistance was prevalent. Multilocus sequence type was generally associated with serovar and provided a reliable prediction of the most common Salmonella serovars.