Prevalence, Detection and Identification of Listeria monocytogenes in Retail Chicken Meat in Ludhiana, India by Employing conventional Isolation Techniques and Molecular Polymerase Chain Reaction (PCR) Assay

Arcobacters are food and waterborne pathogens associated with human and animal infections. The objective of the present study was to investigate the prevalence and diversity of Arcobacter spp. in commercially sold chicken meat in İzmir region of Turkey. For this purpose, 100 samples including legs (n=40), 17 chicken quarters (n=17), drumstickers (n=16), breasts (n=11), wings (n=10), and carcasses (n=6) were collected from different retail markets. A total of 65 isolates were confirmed as Arcobacter spp. from 55 samples by genus-specific polymerase chain reaction (PCR). The prevalence of Arcobacter spp. was 32.5, 81.3, 64.7, 72.7, 83.3, and 50% for legs, drumstickers, chicken quarters, breasts, carcasses and wings, respectively. Based on the multiplex-PCR, most of the isolates were identified as A. butzleri (n=45, 80%), followed by A. cryaerophilus (n=2, 3.6%), A. skirrowii (n=1, 1.8%) and 17 isolates (30.9%) could not be identified at the species level.

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Application of a Multiplex Polymerase Chain Reaction Assay for the Simultaneous Confirmation of Listeria monocytogenes and Other Listeria Species in Turkey Sample Surveillance†

Journal of Food Protection ◽

10.4315/0362-028x-65.5.780 ◽

2002 ◽

Vol 65 (5) ◽

pp. 780-785 ◽

Cited By ~ 30

Author(s):

IRENE V. WESLEY ◽

KAREN M. HARMON ◽

JAMES S. DICKSON ◽

ANN RAMOS SCHWARTZ

Keyword(s):

Polymerase Chain Reaction ◽

Listeria Monocytogenes ◽

Multiplex Pcr ◽

Multiplex Polymerase Chain Reaction ◽

Rrna Gene ◽

Chain Reaction ◽

Pcr Assay ◽

Listeria Species ◽

Multiplex Pcr Assay ◽

Polymerase Chain

A multiplex polymerase chain reaction was developed to simultaneously identify Listeria monocytogenes and species of the genus Listeria. Two sets of primers were used, with the first amplifying a 938-bp region of the 16S rRNA gene that is highly conserved in all Listeria species and the second amplifying a 174-bp region of the listeriolysin (hlyA) gene of L. monocytogenes. Thus, isolates of Listeria spp. yield a single 938-bp product, whereas L. monocytogenes isolates yield both the 938-bp product and a 174-bp product. The specificity of the assay was verified with all six Listeria species and 11 serotypes of L. monocytogenes, as well as nonrelated bacteria. The multiplex PCR assay was used to determine the incidence of Listeria spp., especially L. monocytogenes, in mechanically separated turkey samples (n = 150 samples). L. monocytogenes strains were selected by using the University of Vermont two-step enrichment protocol and plating to selective Palcam agar. The multiplex PCR assay was used for verification of presumptive Listeria colonies. Approximately 38% of mechanically separated turkey samples (57 of 150) yielded L. monocytogenes; an additional 18% of these samples (27 of 150) harbored other Listeria spp. Fifty-one percent (29 of 57) of the L. monocytogenes isolates were of serogroup 1, 44% (25 of 57) were of serogroup 4, and 2% (1 of 57) were assigned to serogroups other than 1 and 4.

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Assessing the Microbial Quality (Listeria monocytogenes) of Chicken Meat by Polymerase Chain Reaction in Different Areas of Chennai

International Journal of Current Microbiology and Applied Sciences ◽

10.20546/ijcmas.2018.705.114 ◽

2018 ◽

Vol 7 (05) ◽

pp. 932-935

Author(s):

S. Satheesh Raja ◽

V. Apparao ◽

R. Narendra Babu ◽

M. Thamizhannal

Keyword(s):

Polymerase Chain Reaction ◽

Listeria Monocytogenes ◽

Chicken Meat ◽

Microbial Quality ◽

Chain Reaction ◽

Polymerase Chain

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Polymerase Chain Reaction Detection of Listeria monocytogenes on Frankfurters Using Oligonucleotide Primers Targeting the Genes Encoding Internalin AB

Journal of Food Protection ◽

10.4315/0362-028x-66.2.237 ◽

2003 ◽

Vol 66 (2) ◽

pp. 237-241 ◽

Cited By ~ 36

Author(s):

YONG SOO JUNG ◽

JOSEPH F. FRANK ◽

ROBERT E. BRACKETT ◽

JINRU CHEN

Keyword(s):

Polymerase Chain Reaction ◽

Listeria Monocytogenes ◽

Meat Products ◽

Chain Reaction ◽

Pcr Assay ◽

Oligonucleotide Primers ◽

Pcr Product ◽

Genes Encoding ◽

Pure Cell ◽

Polymerase Chain

A polymerase chain reaction (PCR) assay targeting the genes encoding internalin AB (inlAB) was developed for detecting Listeria monocytogenes in pure cell cultures and on artificially contaminated frankfurters. Four sets of oligonucleotide primers were evaluated. The set targeting a 902-bp region of the inlAB gene was the most specific. This PCR product was detected in 51 L. monocytogenes strains belonging to four different serogroups (1/2a, 1/2b, 1/2c, and 4b). In contrast, the PCR product was not detected in other Listeria spp. (Listeria innocua, Listeria ivanovii, Listeria seeligeri, Listeria welshimeri, or Listeria grayi) or in gram-positive, non-Listeria bacteria, indicating that the primer set was highly specific for L. monocytogenes. The detection limit of the PCR assay was 105 CFU per ml of pure cell culture. However, the assay could detect as few as 101 CFU of L. monocytogenes in 25 g of frankfurter with 16 h of enrichment in modified Listeria enrichment broth at 30°C. The total assay time including enrichment was approximately 24 h. These results suggest that the PCR assay can be used to rapidly detect L. monocytogenes on frankfurters and possibly other types of ready-to-eat meat products.

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Incidence and Polymerase Chain Reaction Assay of Listeria monocytogenes from Raw Milk in Gyeongnam Province of Korea

Journal of Food Protection ◽

10.4315/0362-028x-65.1.111 ◽

2002 ◽

Vol 65 (1) ◽

pp. 111-115 ◽

Cited By ~ 5

Author(s):

KWANG-SOO HA ◽

SEON-JA PARK ◽

SOOK-JAE SEO ◽

JUNG-HYUN PARK ◽

DUCK-HWA CHUNG

Keyword(s):

Polymerase Chain Reaction ◽

Listeria Monocytogenes ◽

Raw Milk ◽

Food Samples ◽

Chain Reaction ◽

Pcr Assay ◽

Biochemical Tests ◽

Milk Samples ◽

Polymerase Chain ◽

Pcr Assays

A total of 50 raw milk samples from Gyeongnam Province of Korea were examined for the incidence of Listeria monocytogenes between July 1998 and August 1998. L. monocytogenes isolated by biochemical test was confirmed by polymerase chain reaction (PCR) with two sets of primers designed from the invasion-associated protein (iap) gene. After standard PCR with external primers, the amplified DNA was confirmed by a second round of PCR with internal primers (nested PCR). Both the external and internal primers generated 468-bp and 287-bp products, respectively. Only one (G9 strain) of the three suspect samples that tested positive in biochemical tests for L. monocytogenes from 50 raw milk samples was also PCR positive. Following this procedure, PCR-positive G9 strain was confirmed by Southern blot using the 287-bp internal iap probe again. The detection limit of G9 strain by standard PCR assay was as few as 102 cells, equivalent to approximately 1 pg of L. monocytogenes DNA. These PCR assays may be useful for novel detection as well as rapid confirmation for L. monocytogenes from food samples and the field.

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Polymerase chain reaction amplification and typing of rotavirus in environmental samples

Water Science & Technology ◽

10.2166/wst.1995.0643 ◽

1995 ◽

Vol 31 (5-6) ◽

pp. 371-374 ◽

Cited By ~ 5

Author(s):

R. Gajardo ◽

R. M. Pintó ◽

A. Bosch

Keyword(s):

Polymerase Chain Reaction ◽

Environmental Samples ◽

Polymerase Chain Reaction Amplification ◽

Rt Pcr ◽

Chain Reaction ◽

Pcr Assay ◽

Group A ◽

Reaction Amplification ◽

Stool Specimens ◽

Polymerase Chain

A reverse transcription polymerase chain reaction (RT-PCR) assay is described that has been developed for the detection and serotyping of group A rotavirus in stool specimens and concentrated and non-concentrated sewage specimens.

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Analysis and validation of a highly sensitive one-step nested quantitative real-time polymerase chain reaction assay for specific detection of severe acute respiratory syndrome coronavirus 2

Virology Journal ◽

10.1186/s12985-020-01467-y ◽

2020 ◽

Vol 17 (1) ◽

Author(s):

Yang Zhang ◽

Chunyang Dai ◽

Huiyan Wang ◽

Yong Gao ◽

Tuantuan Li ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Real Time ◽

Quantitative Polymerase Chain Reaction ◽

Clinical Samples ◽

Chain Reaction ◽

Pcr Assay ◽

Qrt Pcr ◽

Highly Sensitive ◽

One Step ◽

Polymerase Chain

Abstract Background Coronavirus disease 2019 (COVID-19), caused by SARS-CoV-2, is posing a serious threat to global public health. Reverse transcriptase real-time quantitative polymerase chain reaction (qRT-PCR) is widely used as the gold standard for clinical detection of SARS-CoV-2. Due to technical limitations, the reported positive rates of qRT-PCR assay of throat swab samples vary from 30 to 60%. Therefore, the evaluation of alternative strategies to overcome the limitations of qRT-PCR is required. A previous study reported that one-step nested (OSN)-qRT-PCR revealed better suitability for detecting SARS-CoV-2. However, information on the analytical performance of OSN-qRT-PCR is insufficient. Method In this study, we aimed to analyze OSN-qRT-PCR by comparing it with droplet digital PCR (ddPCR) and qRT-PCR by using a dilution series of SARS-CoV-2 pseudoviral RNA and a quality assessment panel. The clinical performance of OSN-qRT-PCR was also validated and compared with ddPCR and qRT-PCR using specimens from COVID-19 patients. Result The limit of detection (copies/ml) of qRT-PCR, ddPCR, and OSN-qRT-PCR were 520.1 (95% CI: 363.23–1145.69) for ORF1ab and 528.1 (95% CI: 347.7–1248.7) for N, 401.8 (95% CI: 284.8–938.3) for ORF1ab and 336.8 (95% CI: 244.6–792.5) for N, and 194.74 (95% CI: 139.7–430.9) for ORF1ab and 189.1 (95% CI: 130.9–433.9) for N, respectively. Of the 34 clinical samples from COVID-19 patients, the positive rates of OSN-qRT-PCR, ddPCR, and qRT-PCR were 82.35% (28/34), 67.65% (23/34), and 58.82% (20/34), respectively. Conclusion In conclusion, the highly sensitive and specific OSN-qRT-PCR assay is superior to ddPCR and qRT-PCR assays, showing great potential as a technique for detection of SARS-CoV-2 in patients with low viral loads.

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