Detection of hilA gene sequences in serovars of Salmonella enterica sufigbspecies enterica

Cardona-Castro, Nora; Restrepo-Pineda, Eliana; Correa, Margarita M.

doi:10.1590/s0074-02762002000800016

Cited by 42 publications

(35 citation statements)

References 16 publications

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“…The hilA gene is an important feature of Salmonella pathogenesis, as it is required for bacterial colonization of the extracellular, luminal compartment of the host intestine (Murray & Lee, 2000). Hence, in the present study, the specificity and sensitivity of a pair of primers targeting the hilA gene of Salmonella serovars (Cardona-Castro et al, 2002) were assessed for the detection of Salmonella species in human faeces. The primers were originally designed with the intention of cloning an hilA gene fragment in order to disrupt this gene in the chromosome and elucidate its function in Salmonella species that are clinically important for humans (unpublished results).…”

Section: Resultsmentioning

confidence: 99%

“…A 30-bp forward primer (59-CGGAACGTTATTTGCGCCATGCTGAGGTAG-39) and a 27-bp reverse primer (59-GCATGGATCCCCGCCGGCGAGATTGTG-39) (Cardona-Castro et al, 2002), targeting the hilA gene of Salmonella Typhimurium, were used in PCR to obtain a 784-bp product. Amplification was carried out in a total volume of 50 ìl containing 25 pmol each primer, 50 ìM each dNTP, 3 mM MgCl 2 , 1·5 U Taq DNA polymerase, 13 PCR buffer and 4 ìl template.…”

Section: Methodsmentioning

confidence: 99%

“…As Salmonella infection is the leading food-borne disease in Malaysia (Yasin et al, 1995), we were interested to test a variety of primers to improve target specificity. In the present study, we have evaluated the specificity of a pair of primers reported by Cardona-Castro et al (2002) that targets the hilA gene of Salmonella strains. The method presented here is rapid, simple, specific and sensitive, as Salmonella serovars can be detected directly from faecal samples with no prior extraction of genomic DNA.…”

Section: Introductionmentioning

confidence: 99%

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Simple and rapid detection of Salmonella strains by direct PCR amplification of the hilA gene

Pathmanathan

Cardona-Castro

Sánchez-Jiménez

et al. 2003

Journal of Medical Microbiology

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Simple and rapid detection of Salmonella strains by direct PCR amplification of the hilA gene

Pathmanathan

Cardona-Castro

Sánchez-Jiménez

et al. 2003

Journal of Medical Microbiology

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“…Secondly, PCR amplifications were conducted using two primer sets targeting sefC and fliC genes for confirmation of both S. Enteritidis and S. Typhimurium serovars, respectively, according to the procedures reported previously [18,19]. Moreover, five specific primer pairs were used for PCR detection of various virulence genes located on SPIs and plasmids (hilA, sopB, stn, pefA, and spvC genes) based on the protocols of several investigators [17,18,20,21]. The primer sequences and their corresponding genes are listed in Table 1.…”

Section: Virulence Genotyping Of Salmonella Isolates By Polymerase Chmentioning

confidence: 99%

Virulence genotypes of clinical SalmonellaSerovars from broilers in Egypt

Ammar

Mohamed

El-Hamid

et al. 2016

J Infect Dev Ctries

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Introduction: Salmonella serovars are one of the primary foodborne pathogens. Poultry consumption is responsible for the majority of disease cases worldwide. The prevalence of virulence determinants among Salmonella serovars appears to be lacking in Egypt. Therefore, this study investigated the occurrence, antibiotic resistance patterns, and virulence gene profiling of Salmonella serovars in broilers. Methodology: Three hundred samples from broiler chickens were examined for the presence of Salmonella by standard microbiological techniques. All Salmonella isolates were tested for their sensitivity against ten antibiotics and subjected to virulence genotyping by polymerase chain reaction (PCR). Results: The overall isolation percentage of Salmonella was 17%. Seven different serovars were found, with the main one being Salmonella Typhimurium (52.94%). Salmonella isolates were sensitive to most of the tested antibiotics, but they exhibited absolute resistance against amoxicillin/clavulanic acid. Nine Salmonella strains (52.94%) were resistant to at least three antibiotics. Further PCR investigations into 17 Salmonella strains revealed different distribution patterns of eight virulence determinants among the isolates. The invA gene was the most prevalent one (100%), followed by hilA (88.24%), stn (58.82%), and fliC genes (52.94%), while each of sopB and pefA genes had a similar prevalence (41.18%), and sefC and spvC genes had the lowest prevalence (11.76 and 5.88%, respectively). PCR genotyping allowed grouping of Salmonella strains into ten genetic profiles. Conclusions: These results will help in understanding the spread of virulence genotypes and antibiotic resistance among Salmonella serovars in broilers.

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“…que van a amplificar (12,(15)(16)(17) Los iniciadores que utilizamos en este trabajo fueron diseñados con el objetivo de detectar el gen hilA en serotipos de Salmonella de importancia clínica (13). El primer US 5´-3´ contiene un sitio de restricción BamHI y 17 nucleótidos y el primer DS 3´-5´ contiene un sitio de restricción Hindi y 21 nucleótidos que concuerdan perfectamente con la secuencia del gen hilA.…”

Section: Discussionunclassified

Desarrollo y evaluación de una prueba de reacción en cadena de la polimerasa (PCR), utilizando la secuencia del gen hilA para diagnóstico de fiebre entérica por Salmonella spp.

Sánchez¹,

Cardona-Castro²

2004

biomedica

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Desarrollo y evaluación de una prueba de reacción en cadena de la polimerasa (PCR), utilizando la secuencia del gen hilA para diagnóstico de fiebre entérica por Salmonella spp. El diagnóstico de fiebre entérica por Salmonella spp. se basa en el aislamiento de la bacteria en hemocultivos el cual consume tiempo, no siempre está disponible y tiene poca utilidad en pacientes con tratamiento antibiótico previo. Por consiguiente, se hace necesario el desarrollo de una prueba rápida, sensible y específica para el diagnóstico de fiebre entérica. Salmonella spp. utiliza el gen hilA (componente de la isla de patogenicidad I) para invadir células epiteliales y producir infección. Al usar la secuencia de este gen se diseñó una prueba de PCR para detectar la bacteria en sangre y se evaluó su sensibilidad, especificidad, valor predictivo positivo y valor predictivo negativo, utilizando la metodología prueba de una prueba. La prueba de oro fue el hemocultivo. Se estudiaron 34 individuos con sintomatología de fiebre entérica con aislamiento de Salmonella serotipo Typhi en hemocultivos; 35 individuos con sepsis por otros bacilos Gram negativos aislados de hemocultivo (Klebsiella pneumoniae, 9; Serratia marcescens, 5; Escherichia coli, 4; Pseudomonas aeruginosa, 9; Providencia alcalifaciens, 4, y Enterobacter cloacae, 4) y 150 muestras de sangre de voluntarios asintomáticos. La sensibilidad, especificidad, valor pronóstico positivo y valor pronóstico negativo de la PCR fue del 100%. El número mínimo de UFC/ml que la PCR detecta en sangre es de 10.Palabras clave: PCR, fiebre entérica, gen hilA, Salmonella spp. Development and evaluation of a PCR method for diagnosis of Salmonella enteric fever, based on DNA sequences of the hilA geneTypically, diagnosis of enteric fever due to Salmonella spp. is by bacterial isolation from blood culture; however, the blood culture method is slow, not always available, and not informative in patients with antibiotic treatment. Salmonella spp. uses the hilA gene (component of the pathogenicity island I) to invade epithelial cells and produce infection. Using the hilA gene sequence a PCR test was designed to detect Salmonella in blood samples. The sensitivity (S), specificity (SP), positive predictive value (PPV) and negative predictive value (NPV) of the PCR method were obtained by testing the blood samples from 34 patients with suspected of enteric fever. Presence of S. typhi was confirmed by blood culture. Blood samples were also tested from 35 patients with infections due to other non-Salmonella pathogens, again corroborated by blood culture (Klebsiella pneumoniae, 9; Serratia marcescens, 5; Escherichia coli, 4; Pseudomonas aeruginosa, 9; Providencia alcalifaciens, 4; Enterobacter cloacae, 4). Control samples were obtained from 150 healthy volunteers. The S, SP, PPV and NPV for the PCR method were all 100%. The lowest number of colony forming units/ml detected by PCR in blood samples was 10.

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Detection of hilA gene sequences in serovars of Salmonella enterica sufigbspecies enterica

Cited by 42 publications

References 16 publications

Simple and rapid detection of Salmonella strains by direct PCR amplification of the hilA gene

Simple and rapid detection of Salmonella strains by direct PCR amplification of the hilA gene

Virulence genotypes of clinical SalmonellaSerovars from broilers in Egypt

Desarrollo y evaluación de una prueba de reacción en cadena de la polimerasa (PCR), utilizando la secuencia del gen hilA para diagnóstico de fiebre entérica por Salmonella spp.

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