A PCR protocol using inl gene as a target for specific detection of Listeria monocytogenes

Almeida, Paulo Fernando de; Almeida, Rogéria Comastri de Castro

doi:10.1016/s0956-7135(99)00067-5

Cited by 39 publications

(28 citation statements)

References 20 publications

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“…In the present study, the inlA gene was detected in all L. monocytogenes isolates (100%); these results were similar to those obtained by Liu et (11) . Studies conducted by Poyart et al and Jung et al showed that the inlA gene is species-specific, thereby suggesting their species-wide sequence conservation (12) (13) .…”

supporting

confidence: 92%

Prevalence, and virulence determination of Listeria monocytogenes strains isolated from clinical and non-clinical samples by multiplex polymerase chain reaction

Pournajaf

Rajabnia

Sedighi

et al. 2016

Rev. Soc. Bras. Med. Trop.

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Introduction: This study aimed to determine the prevalence, and virulence factors of Listeria monocytogenes isolated from various samples by multiplex polymerase chain reaction (MPCR). Methods: A total of 617 isolates were obtained and MPCR was employed for detection of the inlA, inlC, and inlJ genes. Results: L. monocytogenes was detected in 46 (7.45%) of the 617 specimens. inlA, inlC, and inlJ were detected in 100%, 76.26%, and 71% isolates, respectively. Conclusions: This study validated MPCR in the analysis and rapid detection of L. monocytogenes. The role of the genes in pathogenesis of the strains can also be affirmed.

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confidence: 92%

Prevalence, and virulence determination of Listeria monocytogenes strains isolated from clinical and non-clinical samples by multiplex polymerase chain reaction

Pournajaf

Rajabnia

Sedighi

et al. 2016

Rev. Soc. Bras. Med. Trop.

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“…Conventional methods for detection of L. monocytogenes involve selective enrichment, followed by subsequent biochemical tests, which are laborious and time-consuming (Almeida and Almeida, 2000). It is also difficult to determine the number of L. monocytogenes due to high levels of competitive microorganisms (Capita et al, 2001 Martín et al, 2004).…”

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confidence: 99%

Listeria monocytogenes in retailed raw chicken meat in Malaysia

et al. 2012

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This study aimed to determine the prevalence Listeria monocytogenes in raw chicken meat samples at hypermarkets and wet markets. Chicken drumsticks, breasts, and thighs were randomly selected. The most probable number (MPN) PCR method was used to quantify the L. monocytogenes in the samples. Listeria monocytogenes was detected in 20% of the samples. Occurrence of L. monocytogenes was highest in breast (42.03%) followed by drumstick (11.27%) and thigh (7.14%). Samples from hypermarkets showed higher occurrence (25.71%) of L. monocytogenes compared with wet markets (14.29%). The density of L. monocytogenes found in samples ranged from <3.0 to 16 MPN·g −1 . The presence of L. monocytogenes in raw chicken meat is unwanted but unpreventable. Thus, further research on the processing method to reduce and eliminate this kind of bacteria in chicken meat before consumption is necessary. The presence of L. monocytogenes in chicken samples suggests the importance of this pathogen in chicken. Thus, more study is needed to find ways to eliminate this pathogen from poultry.

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“…Rapid enumeration of L. monocytogenes in biofilms could thus be very useful in the identification of sources of food contamination. DNA-based methods, such as PCR, have been considerably developed to detect food-borne pathogens like L. monocytogenes (1,6,8,21). Real-time PCR has previously been proposed to detect and quantify L. monocytogenes in food products like milk, cheese, and cabbage (10,13,18).…”

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confidence: 99%

Quantitative Detection of Listeria monocytogenes in Biofilms by Real-Time PCR

Guilbaud

Coppet²,

Bourion

et al. 2005

Appl Environ Microbiol

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A quantitative method based on a real-time PCR assay to enumerate Listeria monocytogenes in biofilms was developed. The specificity for L. monocytogenes of primers targeting the listeriolysin gene was demonstrated using a SYBR Green I real-time PCR assay. The number of L. monocytogenes detected growing in biofilms was 6 ؋ 10 2 CFU/cm 2 .In the food industry, Listeria monocytogenes represents an important health risk. The consumption of food products contaminated with this gram-positive bacterium can cause listeriosis, a serious disease with a 30% mortality rate. Raw material can contain L. monocytogenes, but contamination of food products can also occur during processing. The rapid methods currently available for identifying L. monocytogenes are limited by a threshold of approximately 10 5 CFU/ml and consequently require enrichment procedures (22). Rapid quantification of L. monocytogenes is important for many reasons, such as in the Hazard Analysis and Critical Control Point method program to verify critical limits and monitor contamination levels and for quantitative risk assessment. L. monocytogenes, like several food-borne pathogens, can attach to the surfaces in contact with the products, leading to the formation of biofilms (12,14). Adherent bacteria can acquire new physiological properties compared with planktonic cells conferring resistance to disinfectants or sanitizers, leading to hygiene problems, particularly for pathogens that can participate in the recontamination of products (7,19). Rapid enumeration of L. monocytogenes in biofilms could thus be very useful in the identification of sources of food contamination. DNA-based methods, such as PCR, have been considerably developed to detect food-borne pathogens like L. monocytogenes (1,6,8,21). Real-time PCR has previously been proposed to detect and quantify L. monocytogenes in food products like milk, cheese, and cabbage (10, 13, 18). However, to our knowledge, no study has been reported concerning real-time PCR for the detection and quantification of L. monocytogenes in biofilms.The aim of this work was to develop a quantitative method to evaluate the population of L. monocytogenes in artificially made biofilms. The real-time PCR can be used to estimate the number of copies of a target gene in a sample and is reported to be more sensitive than conventional qualitative PCR. Realtime PCR is based on the detection and quantification of a fluorescent reporter, whose emission is directly proportional to the quantity of amplicons generated during the PCR. The fluorescent reporter used in this study was SYBR Green I, a nonspecific double-stranded DNA-binding dye. SYBR Green I has the advantage of not requiring the design of specific probes, and its binding is not affected by potential mutations of the target gene. The specificity and sensitivity of the primers used were determined, and then the effectiveness of four methods of extracting DNA from L. monocytogenes growing in biofilms was evaluated. Finally, the detection and quantification of L. monocytogen...

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A PCR protocol using inl gene as a target for specific detection of Listeria monocytogenes

Cited by 39 publications

References 20 publications

Prevalence, and virulence determination of Listeria monocytogenes strains isolated from clinical and non-clinical samples by multiplex polymerase chain reaction

Prevalence, and virulence determination of Listeria monocytogenes strains isolated from clinical and non-clinical samples by multiplex polymerase chain reaction

Listeria monocytogenes in retailed raw chicken meat in Malaysia

Quantitative Detection of Listeria monocytogenes in Biofilms by Real-Time PCR

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