Polymerase chain reaction (PCR4)-based detection of hly and plc-A genes in Listeria monocytogenes isolated from dairy and meat products in Iran

Lotfollahi, Lida; Pournajaf, Abazar; GholamrezaIrajian,; Nowrouzi, Jamileh

doi:10.5897/ajmr2013.6468

Cited by 10 publications

(3 citation statements)

References 14 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Table 1: Preverance of l, monocytogenes in the Market, Buffalo and Cow milk according to conventional method. (Lida et al, 2014 andAnonymous, 2021).…”

Section: Resultsmentioning

confidence: 99%

Phenotypic and Genotypic Characterization of Listeria Monocytogenes Isolated From Raw Milk in Assiut City

GERGIS,

HAFEZ,

MOHAMMED

et al. 2024

Assiut Veterinary Medical Journal

View full text Add to dashboard Cite

Among a food-associated pathogen, Listeria monocytogenes has become the most important one. It can cause a "high fatality rates 20-30%" in comprasion to another foodborne bacteria which causing dangerous disease called listeriosis. Listeria outbreaks are often linked to dairy products and milk. This article intends to review phenotypic and genotypic characters of L. monocytogenes in Assiut city. How common L. monocytogenes is determined in different sources of raw milk (20 market milk, 40 buffleo milk and 40 cow milk). By detection of 3 virulence genes and its sequencing (hlyA, inIB and (prfA) can deterimined the pathogenic potential of the isolates. Firstly enrichment samples in fraser broth, then, plating on to ALOA®, Merck. Finally a multiplex-PCR,was used for identification of suspected colonies. The results revealed that 8 (40%), 6 (15%) and 11 (27%) of samples that collected from market, buffalo and cow raw milk were positive respectively. Antibacteerial sensitive test showing in vitro thai Listeria mmonocytogenes had a higher sensitivity to Sulfa methoxazoletrimthoprim (SXT) and Ciprofloxacin, (CIP) followed by Chloramphenicol (C), low sensitive to Gentamicin (CN), it resisted Erythrommycin (E) and Amoxicillin (AX). The PCR results for isolates have hly A Inl B and prf A genes of L. monoctogenes.

show abstract

“…Table 1: Preverance of l, monocytogenes in the Market, Buffalo and Cow milk according to conventional method. (Lida et al, 2014 andAnonymous, 2021).…”

Section: Resultsmentioning

confidence: 99%

Phenotypic and Genotypic Characterization of Listeria Monocytogenes Isolated From Raw Milk in Assiut City

GERGIS,

HAFEZ,

MOHAMMED

et al. 2024

Assiut Veterinary Medical Journal

View full text Add to dashboard Cite

show abstract

“…RPA assay for detection of L. monocytogenes RPA primers were designed to detect the plcA gene of L. monocytogenes. The target region is an important virulence gene that has been shown to have high specificity for diagnosing L. monocytogenes strain (Lida et al, 2014). The RPA reactions were performed using 3.1 ng of the genomic DNA of L. monocytogenes as a template.…”

Section: Resultsmentioning

confidence: 99%

DIRECT DETECTION OF LISTERIA MONOCYTOGENES BY RECOMBINASE POLYMERASE AMPLIFICATION

Phung

2021

Preprint

View full text Add to dashboard Cite

Listeria monocytogenes is one of the most common types of food poisoning bacteria which can cause serious foodborne diseases or even lethality. Generally, L. monocytogenes can be detected using traditional microbiology or molecular biology techniques, notably PCR. However, the application of these methods at the field is restricted due to the strict requirement of equipment and skilled personnel. In this study, recombinase polymerase amplification (RPA), an isothermal PCR assay was developed to rapidly detect L. monocytogenes in the crude samples. The results showed that the RPA reaction, without requiring complex thermal cycles, was well-performed in the optimal conditions of 39°C within only 25 minutes. The limit of detection was identified as 310 fg of L. monocytogenes genomic DNA, which was 1000-fold more sensitive than the conventional PCR. In addition, RPA also succeeded to directly detect L. monocytogenes cells at a concentration as low as 2.5×101 Colony Forming Unit (CFU)/ml in pure cultures and 2.5×102 CFU/ml in crude samples without sample extraction or processing. Therefore, RPA established in this study could be an alternative standard method to confirm the presence of L. monocytogenes in food. Accordingly, this rapid and sensitive method could be further applied to clinical testing for the diagnosis of L. monocytogenes infection, especially in areas with limited settings.

show abstract

“…The RPA primers targeting the plcA gene of L. monocytogenes (from 204624 to 205577 of the Genebank accession number NC_003210.1) were designed according to the guidelines provided by TwistDx (Cambridge, UK) (https://www.twistdx.co.uk/en/support/rpa-assay-design-2). The target region is an important virulence gene that has been shown to have high specificity for diagnosing L. monocytogenes strain (Lida et al, 2014). The primer set was chosen by assessing the specificity using NCBI BLAST and the amplification effects were also evaluated practically.…”

Section: Primer Designmentioning

confidence: 99%

Direct Recombinase Polymerase Amplification Assay for Accurate and Rapid Detection of Listeria Monocytogenes in Food

Tran

Pham

et al. 2022

J microb biotech food sci

View full text Add to dashboard Cite

Listeria monocytogenes is one of the most common types of food poisoning bacteria which can cause serious foodborne diseases or even lethality. Generally, L. monocytogenes can be detected using traditional microbiology or molecular biology techniques, notably PCR. However, the application of these methods at the field is restricted due to the strict requirement of equipment and skilled personnel. In this study, recombinase polymerase amplification (RPA), an isothermal PCR assay was developed to rapidly detect L. monocytogenes in crude samples. The results showed that the RPA reaction, without requiring complex thermal cycles, was well-performed in the optimal conditions of 39°C within only 25 minutes. The limit of detection was identified as 310 fg of L. monocytogenes genomic DNA, which was 1000-fold more sensitive than the conventional PCR. RPA also succeeded to directly detect L. monocytogenes cells at a concentration as low as 2.5 × 101 Colony Forming Unit (CFU)/mL in pure cultures. In addition, RPA could accurately detect L. monocytogenes at 2.5 × 102 CFU/mL in milk without sample extraction or processing. Therefore, RPA established in this study could be an alternative standard method to confirm the presence of L. monocytogenes in food. Accordingly, this rapid and sensitive method could be further applied to clinical testing for the diagnosis of L. monocytogenes infection, especially in areas with limited settings.

show abstract

Polymerase chain reaction (PCR4)-based detection of hly and plc-A genes in Listeria monocytogenes isolated from dairy and meat products in Iran

Cited by 10 publications

References 14 publications

Phenotypic and Genotypic Characterization of Listeria Monocytogenes Isolated From Raw Milk in Assiut City

Phenotypic and Genotypic Characterization of Listeria Monocytogenes Isolated From Raw Milk in Assiut City

DIRECT DETECTION OF LISTERIA MONOCYTOGENES BY RECOMBINASE POLYMERASE AMPLIFICATION

Direct Recombinase Polymerase Amplification Assay for Accurate and Rapid Detection of Listeria Monocytogenes in Food

Contact Info

Product

Resources

About