Multiplex PCR assay based on the citE2 gene and intergenic sequence for the rapid detection of Salmonella Pullorum in chickens
ABSTRACT: Salmonella is one of the most common Gram-negative pathogens and seriously threatens chicken farms and food safety. This study aimed to establish a multiplex polymerase chain reaction (PCR) approach for the identification of different Salmonella enterica subsp. enterica. The citE2 gene and...
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| Format: | Article |
| Language: | English |
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Elsevier
2022-08-01
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| Series: | Poultry Science |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S0032579122002723 |
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| author | Zhike Liu Yan Yu Tetiana Fotina Roman Petrov Zhanna Klishchova Anatoliy Fotin Jinyou Ma |
| author_facet | Zhike Liu Yan Yu Tetiana Fotina Roman Petrov Zhanna Klishchova Anatoliy Fotin Jinyou Ma |
| author_sort | Zhike Liu |
| collection | DOAJ |
| description | ABSTRACT: Salmonella is one of the most common Gram-negative pathogens and seriously threatens chicken farms and food safety. This study aimed to establish a multiplex polymerase chain reaction (PCR) approach for the identification of different Salmonella enterica subsp. enterica. The citE2 gene and interval sequence of SPS4_00301–SPS4_00311 existed in all S. enterica subsp. enterica serovars by genomic comparison. By contrast, a 76 bp deletion in citE2 was found only in Salmonella Pullorum. Two pairs of special primers designed from citE2 and interval sequence were used to establish the multiplex PCR system. The optimized multiplex PCR system could distinguish Salmonella Pullorum and non-Salmonella Pullorum. The sensitivity of the optimized multiplex PCR system could be as low as 6.25 pg/μL and 104 colony-forming units (CFU)/mL for genomic DNA and Salmonella Pullorum cells, respectively. The developed multiplex PCR assay distinguished Salmonella Pullorum from 33 different Salmonella enterica subsp. enterica serotypes and 13 non-target species. The detection of egg samples artificially contaminated with Salmonella Pullorum, Salmonella Enteritidis, and naturally contaminated 69 anal swab samples showed that results were consistent with the culture method. These features indicated that the developed multiplex PCR system had high sensitivity and specificity and could be used for the accurate detection of Salmonella Pullorum in clinical samples. |
| first_indexed | 2024-04-13T11:43:20Z |
| format | Article |
| id | doaj.art-ebfebf61dfb4453eb9cf4d0cf7efb56a |
| institution | Directory Open Access Journal |
| issn | 0032-5791 |
| language | English |
| last_indexed | 2024-04-13T11:43:20Z |
| publishDate | 2022-08-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Poultry Science |
| spelling | doaj.art-ebfebf61dfb4453eb9cf4d0cf7efb56a2022-12-22T02:48:15ZengElsevierPoultry Science0032-57912022-08-011018101981Multiplex PCR assay based on the citE2 gene and intergenic sequence for the rapid detection of Salmonella Pullorum in chickensZhike Liu0Yan Yu1Tetiana Fotina2Roman Petrov3Zhanna Klishchova4Anatoliy Fotin5Jinyou Ma6College of Animal Science and Technology, Henan Institute of Science and Technology, Xinxiang, Henan 453003, China; Faculty of Veterinary Medicine, Sumy National Agrarian University, Sumy, 40021, UkraineCollege of Animal Science and Technology, Henan Institute of Science and Technology, Xinxiang, Henan 453003, ChinaFaculty of Veterinary Medicine, Sumy National Agrarian University, Sumy, 40021, UkraineFaculty of Veterinary Medicine, Sumy National Agrarian University, Sumy, 40021, UkraineFaculty of Veterinary Medicine, Sumy National Agrarian University, Sumy, 40021, UkraineFaculty of Veterinary Medicine, Sumy National Agrarian University, Sumy, 40021, UkraineCollege of Animal Science and Technology, Henan Institute of Science and Technology, Xinxiang, Henan 453003, China; Corresponding authors: Jinyou MaABSTRACT: Salmonella is one of the most common Gram-negative pathogens and seriously threatens chicken farms and food safety. This study aimed to establish a multiplex polymerase chain reaction (PCR) approach for the identification of different Salmonella enterica subsp. enterica. The citE2 gene and interval sequence of SPS4_00301–SPS4_00311 existed in all S. enterica subsp. enterica serovars by genomic comparison. By contrast, a 76 bp deletion in citE2 was found only in Salmonella Pullorum. Two pairs of special primers designed from citE2 and interval sequence were used to establish the multiplex PCR system. The optimized multiplex PCR system could distinguish Salmonella Pullorum and non-Salmonella Pullorum. The sensitivity of the optimized multiplex PCR system could be as low as 6.25 pg/μL and 104 colony-forming units (CFU)/mL for genomic DNA and Salmonella Pullorum cells, respectively. The developed multiplex PCR assay distinguished Salmonella Pullorum from 33 different Salmonella enterica subsp. enterica serotypes and 13 non-target species. The detection of egg samples artificially contaminated with Salmonella Pullorum, Salmonella Enteritidis, and naturally contaminated 69 anal swab samples showed that results were consistent with the culture method. These features indicated that the developed multiplex PCR system had high sensitivity and specificity and could be used for the accurate detection of Salmonella Pullorum in clinical samples.http://www.sciencedirect.com/science/article/pii/S0032579122002723Salmonella PullorumSalmonella Enteritidismultiplex PCRcitE2interval sequence |
| spellingShingle | Zhike Liu Yan Yu Tetiana Fotina Roman Petrov Zhanna Klishchova Anatoliy Fotin Jinyou Ma Multiplex PCR assay based on the citE2 gene and intergenic sequence for the rapid detection of Salmonella Pullorum in chickens Poultry Science Salmonella Pullorum Salmonella Enteritidis multiplex PCR citE2 interval sequence |
| title | Multiplex PCR assay based on the citE2 gene and intergenic sequence for the rapid detection of Salmonella Pullorum in chickens |
| title_full | Multiplex PCR assay based on the citE2 gene and intergenic sequence for the rapid detection of Salmonella Pullorum in chickens |
| title_fullStr | Multiplex PCR assay based on the citE2 gene and intergenic sequence for the rapid detection of Salmonella Pullorum in chickens |
| title_full_unstemmed | Multiplex PCR assay based on the citE2 gene and intergenic sequence for the rapid detection of Salmonella Pullorum in chickens |
| title_short | Multiplex PCR assay based on the citE2 gene and intergenic sequence for the rapid detection of Salmonella Pullorum in chickens |
| title_sort | multiplex pcr assay based on the cite2 gene and intergenic sequence for the rapid detection of salmonella pullorum in chickens |
| topic | Salmonella Pullorum Salmonella Enteritidis multiplex PCR citE2 interval sequence |
| url | http://www.sciencedirect.com/science/article/pii/S0032579122002723 |
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