Differential diagnosis of Salmonella gallinarum and S. pullorum using PCR-RELP

Park, Myeong-Kyu; Choi, Kwang Shik; Kim, Myeong Chul; Chae, Joon

doi:10.4142/jvs.2001.2.3.213

Cited by 14 publications

(9 citation statements)

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“…In contrast, molecular tests have been applied to accelerate differentiation. 9,14,18 However, they were based on molecular markers identified before the complete genomic sequences for these 479361V DIXXX10.1177/1040638713479361Batis ta et alratA gene for differentiating S. Gallinarum and S. Pullorum From the Faculty of Agriculture and Veterinary Sciences, Universidade biovars were available. Some of the published tests require enzymatic digestion of amplicons 10 or more than 1 polymerase chain reaction (PCR).…”

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confidence: 99%

Polymerase chain reaction assay based on ratA gene allows differentiation between Salmonella enterica subsp. enterica serovar Gallinarum biovars Gallinarum and Pullorum

et al. 2013

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Salmonella Pullorum and Salmonella Gallinarum are classified as biovars of Salmonella enterica subsp. enterica serovar Gallinarum. These salmonellae are the causative agents of Pullorum disease and fowl typhoid, respectively, and are widely distributed throughout the world. Although many developed countries have eradicated these diseases from commercial poultry, they are still the cause of significant economic loss in developing countries. When serovar Gallinarum is isolated, it is difficult to immediately differentiate between biovars because they are antigenically identical by serotyping. However, they cause distinct diseases with different epidemiology, and therefore it is important to differentiate them. This may be done biochemically but takes 2 to 3 days. In the present study, S. Pullorum and S. Gallinarum whole genomes were compared, and 1 genomic region of difference, which is part of the ratA gene, was chosen as a molecular marker for a polymerase chain reaction assay to differentiate rapidly between these organisms. In all, 26 strains of S. Gallinarum and 17 S. Pullorum strains were tested and successfully differentiated by the assay.

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confidence: 99%

Polymerase chain reaction assay based on ratA gene allows differentiation between Salmonella enterica subsp. enterica serovar Gallinarum biovars Gallinarum and Pullorum

et al. 2013

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“…After that, the suspended culture in saline was incubated at 100 8C for 2 h for O antigen and then both O and H antigens suspension centrifuged at 6440)g for 20 min. Finally, the cell suspension was adjusted to be 2)10 9 cfu/ml in saline using the McFarland standard 9 (Adriana et al, 2007;Park et al, 2001).…”

Section: O and H Antigens Preparationmentioning

confidence: 99%

“…The test has been used very extensively in the serodiagnosis of typhoid fever and, in developing countries, remains the only practical test available (Kulkarni & Rego, 1994;Narayanappa, Srıpath, Jagdıshkumar, & Rajanı, 2009;Pokhrel, Karmacharya, Mishra, & Koirala, 2009). For many years the test of choice in diagnosis of Pullorum disease and fowl typhoid has been the slide agglutination which was originally developed by Runnels, Coon, Farley, and Thorp (1927) for use with serum and adapted by Schaffer, MacDonald, Hall, and Bunye (1931) for whole blood by using stained antigens (Park, Choi, Kim, & Chae, 2001). Moreover, in recent years the production of antibodies in laboratory animals has become an essential part of many research projects.…”

Section: Introductionmentioning

confidence: 99%

Prevention and identification ofSalmonellaEnteritidis infection via novel diagnostic stained antigens and polyclonal antibodies

Nalbantsoy

2013

Food and Agricultural Immunology

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Analysis of Salmonella Enteritidis with specific antisera and stained antigen helps to reduce the identification time of the infection and offers clinical and epidemiological advantages both for poultry health and food industry for the control of salmonellosis. In this study, S. Enteritidis O and H specific antisera were obtained from rabbits immunised subcutaneously using formalintreated whole bacterial cells as antigens. S. Enteritidis specific stained O and H antigens were prepared and tested for their diagnostic usefulness. In order to eliminate cross reactivity, Salmonella Adeyo was used as an antigen for the production of H specific polyclonal antibody and stained H antigen. Enzymelinked immunosorbent assay results of antisera were obtained, ranging from 273,30090.041Á312,20090.028 A492 nm. Stained O and H antigen preparations were achieved with a stronger binding degree to the hyperimmune rabbit sera and without being shown cross-reactivity between O and H antigens.

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“…In order to rapid identification of Salmonella, recently, researchers made an effort to investigate simple and applicable molecular methods. Monoplex and Multiplex-PCR, PCR-Restriction Fragment Length Polymorphism (PCR-RFLP), PCR-Single-Strand Conformation Polymorphism (PCR-SSCP), Prophage-specific-PCR, Pulsed Field Gel Electrophoresis (PFGE), IS200 fingerprinting, and other molecular methods were employed to discriminate between serovars of salmonella (Dauga et al 1998;Essam Soliman et al 2009;Mortimer et al 2004;Park et al 2001;Shanmugasundaram et al 2009;Rubino et al 1998;Nair et al 1994;Rychlık et al 2008). Conserved region of rfbJ (encodes O antigen), fliC (encodes phase-1 flagellin) and fljB (encodes phase-2 flagellin) genes were mainly used in the mentioned methods (Dauga et al 1998;Hong et al 2008;Lim et al 2003).…”

Section: Introductionmentioning

confidence: 99%

The analysis of groEL gene in Salmonella enterica subspecies enterica serovar Typhimurium isolated from avians by PCR-Restriction Fragment Length Polymorphism method

et al. 2011

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Salmonella enterica subspecies enterica serovar Typhimurium causes food-borne outbreaks and systemic diseases in humans and animals. groEL gene (also known as mopA gene in S. Typhimurium), possessing conserved sequence, plays an important role in invasion of bacteria. The purpose of present study was to identify the polymorphism of groEL gene among different avians in different regions by PCR-RFLP method. Fifty two S. Typhimurium isolates (Broiler (n = 13), Layer (n = 12), Duck (n = 5), Goose (n = 5), Sparrow (n = 8), Canary (n = 3), Pigeon (n = 5) and Casco parrot (n = 1). were identified using serotyping as well as multiplex-PCR. Then, amplification of groEL gene performed and amplified products subjected to restriction digestion with BsuRI enzyme. Three RFLP profiles, A, B and C, generated DNA fragments between approximately 100-1,000 bp in size, were observed. The RFLP profile A was observed in 35 (67.3%), profile B in 14 (26.9%) and profile C in 3 (5.77%) of isolates. S. Typhimurium isolates recovered from 13 broilers (two of which profile A, 9 profile B and 2 profile C) and from 8 sparrows (two of which profile A, 5 profile B and 1 profile C) showed all three profiles, but 12 layers and other avians (including Canary (n = 3), Goose (n = 5), Duck (n = 5), Pigeon (n = 5) and Casco parrot (n = 1)) showed profile A. None of these profiles was allotted for a special region. The result of present study showed that S. Typhimurium undergoes genetic mutations in groEL gene under unpleasant milieu in different regions and in different avians. Thus, genetic diversity, despite conserved nature of groEL gene in S. Typhimurium, may exist but it depends on the condition where bacteria have settled. To our knowledge, three RFLP profiles of groEL gene generated by BsuRI restriction enzyme were not reported previously.

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Differential diagnosis of Salmonella gallinarum and S. pullorum using PCR-RELP

Cited by 14 publications

References 0 publications

Polymerase chain reaction assay based on ratA gene allows differentiation between Salmonella enterica subsp. enterica serovar Gallinarum biovars Gallinarum and Pullorum

Polymerase chain reaction assay based on ratA gene allows differentiation between Salmonella enterica subsp. enterica serovar Gallinarum biovars Gallinarum and Pullorum

Prevention and identification ofSalmonellaEnteritidis infection via novel diagnostic stained antigens and polyclonal antibodies

The analysis of groEL gene in Salmonella enterica subspecies enterica serovar Typhimurium isolated from avians by PCR-Restriction Fragment Length Polymorphism method

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