Application of Multiplex Polymerase Chain Reaction to the Detection of Pathogens in Food

Perry, Lynda; Heard, Preciaus; Kane, Michael D.; Kim, Hanyoup; Savikhin, Sergei; Domínguez, Wilfredo; Applegate, Bruce

doi:10.1111/j.1745-4581.2007.00083.x

Cited by 38 publications

(18 citation statements)

References 69 publications

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“…The multiplex-PCR protocols gave the same results as individual protocols. Multiplex PCR represents an important tool for the diagnosis and study of a large number of samples because it allows the simultaneous detection of multiple genes, reducing the amount of reagents used and time required to obtain the results (Markoulatos et al 2002, Perry et al 2007). The individual PCR protocols and multiplex-PCR also performed well in the detection of virulence genes in samples of porcine origin.…”

Section: Discussionmentioning

confidence: 99%

Detection of virulence-associated genes of Pasteurella multocida isolated from cases of fowl cholera by multiplex-PCR

et al. 2013

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The current systems of breeding poultry, based on high population density, increase the risk of spreading pathogens, especially those causing respiratory diseases and those that have more than one host. Fowl Cholera (FC) is one such pathogen, and even though it represents one of several avian diseases that should be considered in the differential diagnosis of notifiable diseases that present with sudden death, the pathogenesis and virulence factors involved in FC are still poorly understood. The objective of this study was to investigate twelve genes related to virulence in 25 samples of Pasteurella multocida isolated from FC cases in the southern region of Brazil through the development of multiplex PCR protocols. The protocols developed were capable of detecting all of the proposed genes. The ompH, oma87, sodC, hgbA, hgbB, exBD-tonB and nanB genes were present in 100% of the samples (25/25), the sodA and nanH genes were present in 96% (24/25), ptfA was present in 92% (23/25), and pfhA was present in 60% (15/25). Gene toxA was not identified in any of the samples studied (0/25). Five different genetic profiles were obtained, of which P1 (negative to toxA) was the most common. We concluded that the multiplex-PCR protocols could be useful tools for rapid and simultaneous detection of virulence genes. Despite the high frequency of the analyzed genes and the fact that all samples belonged to the same subspecies of P. multocida, five genetic profiles were observed, which should be confirmed in a study with a larger number of samples.

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Section: Discussionmentioning

confidence: 99%

Detection of virulence-associated genes of Pasteurella multocida isolated from cases of fowl cholera by multiplex-PCR

et al. 2013

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“…Moreover, multiplex assay protocols allow for the simultaneous detection of different genes, reducing the amount of reagents and the time required to obtain results (Perry et al, 2007).…”

Section: Identification Of the Capsule Type Of Pasteurella Multocida mentioning

confidence: 99%

Identification of the capsule type of Pasteurella multocida isolates from cases of fowl cholera by multiplex PCR and comparison with phenotypic methods

Furian

Borges

Pilatti

et al. 2014

Rev. Bras. Cienc. Avic.

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The ability of Pasteurella multocida to invade and multiply in its host is enhanced by the presence of the capsule, one of the most important virulence factors for this bacterium. Capsular typing methods are often used in epidemiological and pathogenesis studies of this agent. Five different serogroups have been identified based on serological typing. However, such tests are laborious, and agglutination of homologous antiserum may fail. The aim of this study was to develop a multiplex PCR protocol for the identification of the hyaD-hyaC and dcbF genes specific to serogroups A and D, respectively, and to compare these results with those of phenotypic tests for 54 strains isolated from fowl cholera cases in southern Brazil. The kappa coefficient and chisquare statistics were calculated to assess the agreement between the diagnostic methods and to determine the significance of the results, respectively. The multiplex PCR was able to detect the evaluated genes. Forty-nine strains (90.74%) were classified into serogroup A, and only two isolates (3.7%) were not identified as belonging to any of the serogroups analyzed. In contrast, with the phenotypic tests, only 41 strains (75.93%) were classified into serogroup A and 11 samples (20.37%) were unidentifiable. Of the strains analyzed, 70.37% were classified into the same serogroup (A) by both methods, and the kappa coefficient (k = 0.017) indicated poor agreement between the tests. Thus, multiplex PCR is an alternative for P. multocida capsular typing, as it allows the simultaneous and rapid detection of genes and also provides a greater strain-typing capacity.

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“…Posteriormente, o plaqueamento diferencial é realizado em meios de cultura sólidos para a obtenção de colônias isoladas para a identificação bioquímica e sorológica [8] . [9] .O uso de métodos alternativos especialmente os de detecção de múltiplos patógenos é extremamente conveniente, já que permitem a detecção simultânea de microrganismos, com a vantagem de usar poucos materiais e entregar rapidamente os resultados [10] . Estes métodos de detecção múltipla já são reportados na literatura para a identificação de indicadores das condições de higiene, como também importantes pató-genos humanos [11] .…”

Section: Introductionunclassified

“…These methods are described by recognized organizations such as: United States Food and Drug Administration (FDA), Association of Official Analytical Chemists (AOAC), International Organization for Standardization (ISO) and United States Department of Agriculture/Food Safety and Inspection Service (USDA/FSIS) [9] . The use of alternative methods especially multipathogen detection tools is increasingly convenient since it enables the detection of more than one microorganism simultaneously, with the advantage of using few materials and quickly delivering results [10] . These multiple detection methods are already reported in the literature for hygiene indicators and important human pathogens as well [11] .…”

Section: Introductionmentioning

confidence: 99%

Comparison between Listeria monocytogenes single and multipathogen detection methods

Souza¹,

Ferreira²,

Pasquini-Netto³

et al. 2017

BBR

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143 Biomedical and Biopharmaceutical Research J o r n a l d e I n v e s t i g a ç ã o B i o m é d i c a e B i o f a r m a c ê u t i c a Comparison between Listeria monocytogenes single and multipathogen detection methods Comparação entre os métodos de detecção de Listeria monocytogenes simples e multipatógenos AbstractMultipathogen detection methods have been used to provide a broad range of microorganism detection. We aimed to identify the most searched pathogens in L. monocytogenes detection methods and evaluate if there are sensitivity and specificity differences between single and multipathogen methods. A systematic review was performed by including studies carried out in order to detect L. monocytogenes in a broad range of food sample. A total of 2770 records were retrieved, of which 191 were selected. The majority of the studies (104) presented multipathogen detection, other 87 searched for L. monocytogenes specifically. From the studies for multipathogen detection; other Listeria species (19.4%), Salmonella spp. (21.6%), E. coli O157:H7 (13.2%), Staphylococcus spp. (9.7%), E. coli (5.7%) were more frequently reported. Sensitivity and specificity calculations were derived from only 26 studies, because they compared their data with an official method. Sensitivity and specificity values were close to 100%, showing that others characteristics such as time and cost should be considered to evaluate alternative methods in further research. As a conclusion, evidence generated regarding L. monocytogenes identification methods contribute to method improvements and listeriosis control.Keywords: L. monocytogenes, methods, pathogens, sensitivity, specificity. ResumoOs métodos de detecção para múltiplos patógenos têm sido usados para fornecer uma ampla faixa de detecção de microrganismos. Objetivou-se identificar os patógenos mais buscados nos métodos de detecção para L. monocytogenes e avaliar se existem diferenças nas taxas de sensibilidade e especificidade entre os métodos que buscaram identificar somente L. monocytogenes e os para múltiplos patógenos. Uma revisão sistemática foi realizada incluindo estudos desenvolvidos para detectar L. monocytogenes em amostras alimentares.

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Application of Multiplex Polymerase Chain Reaction to the Detection of Pathogens in Food

Cited by 38 publications

References 69 publications

Detection of virulence-associated genes of Pasteurella multocida isolated from cases of fowl cholera by multiplex-PCR

Detection of virulence-associated genes of Pasteurella multocida isolated from cases of fowl cholera by multiplex-PCR

Identification of the capsule type of Pasteurella multocida isolates from cases of fowl cholera by multiplex PCR and comparison with phenotypic methods

Comparison between Listeria monocytogenes single and multipathogen detection methods

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