Early Detection of Typhoid by Polymerase Chain Reaction

Haque, Abdul; Ahmed, Janbaz; Qureshi, Javed Anver

doi:10.5144/0256-4947.1999.337

Cited by 46 publications

(40 citation statements)

References 17 publications

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“…This problem of sensitivity could be circumvented by using the PCR, which can detect very small amounts of DNA by enzymatic amplification with absolute specificity within several hours. For PCR, we preferred to target the flagellin gene, because the flagellin gene of S. Typhi has unique nucleotide sequences in the hyper variable region of the gene [3,16,17,18,19] which are different from those sequences in other strains of Salmonella. Our PCR assay using these primers of flagellin gene was specific for the S. Typhi strain.…”

Section: Discussionmentioning

confidence: 99%

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Comparative Diagnosis of Typhoid Fever by Polymerase Chain Reaction and Widal Test in Southern Districts (Bannu, Lakki Marwat and D.I.KHAN) of Khyber Pakhtunkhwa, Pakistan

Khan¹,

Sajid²,

Javed³

et al. 2017

Acta sci. Malays

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Typhoid fever is a major dilemma in developing nations, despite the antibiotic use and new antibiotics discovery. Blood culture and serological tests (especially Widal test) that are regularly performed in Pakistan for diagnosis provide objectionable levels of false negative and false positive results, respectively. The present study was carried out at Kohat University of science and technology, Kohat, Pakistan from January 2013 to December 2013. A polymerase chain reaction (PCR) assay was compared with Widal test and blood culture for blood samples of 96 patients. In this study, blood from 25 healthy individuals was collected as a negative control. The detection rate of PCR was maximum (64.5%) followed by Widal test (26%) and blood culture (14.5%). Among the 82 (85.4%) samples that were found negative with blood culture, 48 (58.5%) were successfully detected by PCR and 11 (13.4%) were positive for the Widal test. Findings of this study showed that PCR is a fast and reliable technique for diagnosis of typhoid fever in suspected cases, as compared to frequently used conventional techniques like blood culture and Widal test.

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Section: Discussionmentioning

confidence: 99%

“…The widely used diagnostic practices include Widal test and blood culture because these methods are inexpensive while the other methods are invasive, tedious, and time consuming and so expensive [18]. The Widal test has been found nonspecific and difficult to interpret the results in those areas where typhoid fever is endemic [19,20].…”

Section: Introductionmentioning

confidence: 99%

Comparative Diagnosis of Typhoid Fever by Polymerase Chain Reaction and Widal Test in Southern Districts (Bannu, Lakki Marwat and D.I.KHAN) of Khyber Pakhtunkhwa, Pakistan

Khan¹,

Sajid²,

Javed³

et al. 2017

Acta sci. Malays

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“…DNA extraction: Extraction of DNA from whole blood was done as per procedure described by Haque et al, [11] with several modifications. Blood samples collected in EDTA containers inoculated into BHI broth and incubated for 6 h. One ml of supernatant was centrifuged at 10,000 rpm for 5 min.…”

Section: Methodsmentioning

confidence: 99%

Detection of Salmonella in Blood by PCR using iroB gene

Ganesan

Harish

Menezes

et al. 2014

JCDR

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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

“…El método para la extracción del ADN en muestras de los pacientes y en las muestras contaminadas artificialmente fue el de solución tamponada de lisis descrito por Haque et al (12) al que se le introdujeron las modificaciones que se describen a continuación. Brevemente, se realizó la extracción de ADN a 1 ml de la sangre artificialmente contaminada y a 1 ml de la muestra del paciente.…”

Section: Extracción Del Adn Bacterianounclassified

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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Early Detection of Typhoid by Polymerase Chain Reaction

Cited by 46 publications

References 17 publications

Comparative Diagnosis of Typhoid Fever by Polymerase Chain Reaction and Widal Test in Southern Districts (Bannu, Lakki Marwat and D.I.KHAN) of Khyber Pakhtunkhwa, Pakistan

Comparative Diagnosis of Typhoid Fever by Polymerase Chain Reaction and Widal Test in Southern Districts (Bannu, Lakki Marwat and D.I.KHAN) of Khyber Pakhtunkhwa, Pakistan

Detection of Salmonella in Blood by PCR using iroB gene

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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