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

Pathmanathan, Siva Gowri; Cardona-Castro, Nora; Sánchez-Jiménez, Miryan Margot; Correa-Ochoa, Margarita M.; Puthucheary, S. D.; Thong, Kwai Lin

doi:10.1099/jmm.0.05188-0

Cited by 96 publications

(65 citation statements)

References 21 publications

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“…The invA gene of Salmonella contains sequences unique to this genus and has been proved to be a suitable PCR target with potential diagnostic application 18 . This gene is recognized as an international standard for detection of Salmonella genus 19 .…”

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confidence: 99%

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Multiplex-PCR Assay for Detection of some Major Virulence Genes of <i>Salmonella enterica</i> Serovars from Diverse Sources

et al. 2016

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confidence: 99%

“…Spiking of stool sample was performed according to Pathmanathan et al 18 with some modifications. Briefly, 1 g of stool sample from a healthy individual was mixed with 10 ml of BHI broth.…”

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confidence: 99%

Multiplex-PCR Assay for Detection of some Major Virulence Genes of <i>Salmonella enterica</i> Serovars from Diverse Sources

et al. 2016

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“…Estos iniciadores no amplificaron bandas inespecíficas en las muestras de pacientes infectadas con otras enterobacterias; además, han sido probados por otros autores con otras bacterias y no han amplificado secuencias inespecíficas (19).…”

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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“…Further studies have reported that PCR with primers for hilA which are specific for Salmonella spp. are sensitive and specific for detection of Salmonella in many clinical samples (17)(18)(19) as well as carriers (9).…”

Section: Discussionmentioning

confidence: 99%

Molecular Diagnosis of Salmonella enterica and Shigella spp. in Stool Sample of Children With Diarrhea in Tehran

Alizadeh-Hesar¹,

Bakhshi

Peerayeh³

2014

Int J Enteric Pathog

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Background: Diarrheal illnesses caused by Salmonella and Shigella spp remain a serious public health problem in industrializing countries, and are still an important cause of morbidity and mortality in developed as well as undeveloped countries. Rapid detection of agents that cause diarrhea may help prevent occurrence of outbreaks. Objectives: The aim of this study was to compare conventional biochemical tests, namely API 20E strip system and Polymerase Chain Reaction (PCR) for identification of Salmonella enterica and Shigella spp. isolated from stool samples of patients with diarrhea. Materials and Methods: Stool samples were collected from patients with diarrhea during a three-year period (2009)(2010)(2011)(2012). Conventional biochemical tests were used for identification of Shigella spp. and Salmonella enterica isolates. API 20E strips and PCR methods with oligonucleotide primers specific for ipaH of Shigella genus and hilA of Salmonella enterica were used to confirm the identity of the isolates. Results: Of the 81 suspected S. enterica and 112 Shigella spp. identified by conventional biochemical tests, 77 and 105 were identified as S. enterica and Shigella spp. by API 20E, respectively. All of the isolates were assigned to bacterial species with 99.9% probability value. All of the 81 suspected Salmonella enterica isolates produced 784 bp amplification bands of hilA gene. Among the 112 Shigella isolates confirmed by PCR, 90 (80.35%) were positive for wbgZ and 14 (12.5%) were positive for rfc genes indicative of S. sonnei and S. flexneri , respectively. Conclusions: In conclusion, the results of this study suggest that the PCR amplification of hilA and ipaH is a promising method for identification of Salmonella enterica and Shigella spp. The outcomes of this study can help towards more accurate and easy screening of large population of patients with Salmonella enterica and Shigella spp infections.

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

Cited by 96 publications

References 21 publications

Multiplex-PCR Assay for Detection of some Major Virulence Genes of <i>Salmonella enterica</i> Serovars from Diverse Sources

Multiplex-PCR Assay for Detection of some Major Virulence Genes of <i>Salmonella enterica</i> Serovars from Diverse Sources

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.

Molecular Diagnosis of Salmonella enterica and Shigella spp. in Stool Sample of Children With Diarrhea in Tehran

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