Detection of Salmonella DNA in Chicken Embryos and Environmental Samples by Polymerase Chain Reaction.

Tuchili, L. M.; Kodama, Hiroshi; Sharma, Ravindra; Pandey, G. S.; Kabilika, Swithin; Mukamoto, Masafumi; Tsuji, Sachiyo; Baba, Tomoko

doi:10.1292/jvms.58.881

Cited by 9 publications

(11 citation statements)

References 11 publications

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“…These timeconsuming methods should be complemented with a rapid and primary screening procedure such as PCR. In the PCR studies with avian samples for Salmonella, preparation of the sample, PCR, and detection of the amplicon without and with (required for subclinically infected birds) PE may well take up to 6 and 24 h per sample, respectively (26,33). In this study, we combined capillary PCR and capillary gel electrophoresis to more quickly determine if an avian intestinal sample harbors Salmonella.…”

Section: Figmentioning

confidence: 99%

“…PCR methods and DNA extraction procedures for Salmonella have been described for human, animal, and food samples (5,6,21,23,24). However, little information exists about these techniques used with poultry fecal samples (15,26). These methods seem to be expensive and time-consuming (15,26).…”

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confidence: 99%

“…However, little information exists about these techniques used with poultry fecal samples (15,26). These methods seem to be expensive and time-consuming (15,26). This study describes a practical DNA extraction procedure from primary enrichment cultures of Salmonella, genus-specific capillary PCR, and capillary gel electrophoresis to detect chickens carrying the viable Salmonella in their intestines.…”

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confidence: 99%

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Detection of Salmonellae in Chicken Feces by a Combination of Tetrathionate Broth Enrichment, Capillary PCR, and Capillary Gel Electrophoresis

Çarlı¹,

Unal²,

Caner³

et al. 2001

J Clin Microbiol

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This report describes a rapid detection procedure for salmonellae from chicken feces by the combination of tetrathionate primary enrichment (preenrichment [PE])-bacterial lysis-capillary PCR and capillary gel electrophoresis. Pure Salmonella enterica serovar Enteritidis 64K was reisolated and detected by capillary PCR after buffered peptone water and nutrient broth, tetrathionate broth base Hajna (TTBH), and tetrathionate broth (TTB) preenrichments. When the same culture was mixed with intestinal homogenate, bacteriological reisolation and capillary PCR detection was achieved only by TTBH and TTB preenrichments. Capillary gel electrophoresis revealed that a Salmonella genus-specific 281-bp PCR product was detected when Salmonella strains but not non-Salmonella strains were tested. The detection limit of capillary PCR with whole-cell DNA extracted from pure Salmonella enterica serovars Enteritidis 64K, Typhimurium LT2-CIP60-62, and Gallinarum 64K was 3, 3, and 9 CFU ml ؊1 , respectively. The detection limit of capillary PCR from whole-cell DNA extracted from intestinal homogenate artificially contaminated with the same three strains was 3, 3, and 7 CFU ml ؊1 , respectively. We compared the results of the capillary PCR and bacteriological examination from the natural samples. Thirty-five of 53 naturally contaminated samples produced a specific PCR product. In 9 of the 35 PCR-positive samples, Salmonella could not be detected bacteriologically either by PE or a primary and delayed secondary enrichment (DSE) combination. In the 18 PCR-negative samples, 4 samples were found to harbor Salmonella by both PE and DSE and 14 samples were positive after DSE. Fifty-three additional intestinal homogenate samples, which were negative by their PE and DSE in bacteriological examination, were found to be also negative by their PCRs. The total time required to detect Salmonella with the capillary PCR method we used was approximately 20 h. If samples are from clinically diseased birds, the total time for PCR and detection is reduced to 2 h since the 18-h PE is not required. These results indicate that TTB enrichment, bacterial lysis, and genus-specific capillary PCR combined with capillary gel electrophoresis constitute a sensitive and selective procedure which has the potential to rapidly identify Salmonella-infected flocks.Salmonellae are among the major bacterial pathogens of poultry in Turkey and in the world (3,8). Prevention of Salmonella infection is important for poultry health and for the food industry, and prevention can be achieved only by good monitoring and screening programs (10,17,27). Salmonella detection by bacteriological methods usually requires 5 to 11 days (10, 27), and samples with low numbers of Salmonella cells, usually seen in subclinically infected chickens, may give false-negative results (7). Efforts have been made to reduce the time required and to increase the sensitivity of methods to detect Salmonella serovars in poultry samples (16,25). PCR with preincubation in an enrichment broth has been perform...

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Section: Figmentioning

confidence: 99%

mentioning

confidence: 99%

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confidence: 99%

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Detection of Salmonellae in Chicken Feces by a Combination of Tetrathionate Broth Enrichment, Capillary PCR, and Capillary Gel Electrophoresis

Çarlı¹,

Unal²,

Caner³

et al. 2001

J Clin Microbiol

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“…Using PCR, 3%, 8.4% and 6% albumen samples collected from Balady, white and brown eggs respectively were positive for Salmonella serovars. Salmonella DNA could be detected from infertile eggs which incidence was higher than that by bacteria isolation [24]. Salmonella strains grow better in fresh egg white than in egg white of 2 or 3 weeks old [17].…”

Section: Discussionmentioning

confidence: 99%

Multiplex PCR-based detection of Salmonella typhimurium and Salmonella enteritidis in Specific Pathogen Free (SPF) and Commercial Eggs

El-Jakee¹,

Khelfa²,

El-Safty³

et al. 2016

Clin Microbiol

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Salmonella serovars are one of the major bacterial causes of food borne diseases. Eggs are commonly identified as food sources responsible for salmonellosis outbreaks. This study aimed to isolate Salmonella Typhimurium and Salmonella Enteritidis from 1750 hens' eggs, and use of multiplex polymerase chain reaction (Multiplex PCR) in the identification of different Salmonella serovars from eggs. The incidence of salmonellae among the Balady eggs yolk was 1.3%, while the incidence was 1.2% among white and brown eggs samples (each). S. Typhimurium and S. Enteritidis were identified (0.6 and 0.5% respectively). The isolates were confirmed using fliC, sefA genes and gene specific for genus Salmonella. All albumen samples negative for isolation of isolation of salmonellae by culture method were retested by PCR.From the retested albumen samples 3%, 8.4% and 6% collected from Balady, white and brown eggs respectively were positive for Salmonella serovars using Multiplex PCR. No salmonellae could be detected from specific pathogen free (SPF) eggs using both PCR and conventional methods.

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“…izolasyonu yapılamadı. İzolasyon çalışmaları ile ilgili diğer araştırı-cıların bulguları farklılık göstermektedir (8,22,23,32 Sonuç olarak bu çalışmada, kloakal svap alınan 50 kümesin 6'sında (%12) örnek bazında da 100 örneğin 6'sından (%6) selektif zenginleştirmeden sonra yapılan PCR ve kültür metodu ile Salmonella'nın cins düzeyinde varlığı tespit edildi. Yumurta örneklerinden ise her iki metotla da Salmonella belirlenemedi.…”

Section: Tartışma Ve Sonuçunclassified

Ankara Bölgesi’ndeki tavukçuluk işletmelerinden Salmonella spp. izolasyonu

AYDIN¹

2008

Ankara Üniversitesi Veteriner Fakültesi Dergisi

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Detection of Salmonella DNA in Chicken Embryos and Environmental Samples by Polymerase Chain Reaction.

Cited by 9 publications

References 11 publications

Detection of Salmonellae in Chicken Feces by a Combination of Tetrathionate Broth Enrichment, Capillary PCR, and Capillary Gel Electrophoresis

Detection of Salmonellae in Chicken Feces by a Combination of Tetrathionate Broth Enrichment, Capillary PCR, and Capillary Gel Electrophoresis

Multiplex PCR-based detection of Salmonella typhimurium and Salmonella enteritidis in Specific Pathogen Free (SPF) and Commercial Eggs

Ankara Bölgesi’ndeki tavukçuluk işletmelerinden Salmonella spp. izolasyonu

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