Development of a triplex PCR assay for the specific detection of Campylobacter jejuni, Salmonella spp., and Escherichia coli O157:H7

2003 ◽  
Vol 17 (4) ◽  
pp. 135-138 ◽  
Author(s):  
Carl Gilbert ◽  
Debra Winters ◽  
Awilda O'Leary ◽  
Michael Slavik
2020 ◽  
Vol 13 (1) ◽  
pp. 66-80
Author(s):  
Hồ Thị Thanh Thủy ◽  
Nguyễn Văn Trường ◽  
Nguyễn Bảo Toàn ◽  
Lao Đức Thuận ◽  
Trương Kim Phượng ◽  
...  

Ngộ độc thực phẩm, với một trong những nguyên nhân chính do nhiễm khuẩn vẫn luôn là mối lo ngại, mang tính toàn cầu, được Tổ chức Sức khỏe Thế giới rất quan tâm. Việc xác định chính xác đối tượng vi khuẩn nhiễm vẫn luôn là một nhu cầu cấp thiết của các labo lâm sàng. Nghiên cứu trước đây của chúng tôi đã được công bố với việc thành công bước đầu trong việc xây dựng một quy trình dựa trên kĩ thuật PCR-Reverse Dot Blot (PCR-RDB) nhằm xác định đồng thời 12 vi khuẩn gây bệnh đường ruột ch yếu, bao gồm Bacillus cereus, Clostridium botulinum, Clostridium perfringen, Staphylococcus aureus, Listeria monocytogenes, Escherichia coli O157:H7, Salmonella spp., Shigella spp.., Vibrio cholerae, Vibrio parahaemolyticus, Yersinia enterocolitica và Brucella spp. Chúng tôi tiếp tục phát triển nghiên cứu này nhằm hoàn thiện quy trình PCR-RDB bằng việc thiết kế bổ sung trên màng các loại mẫu dò nhằm làm chứng dương, chứng âm, chứng màu và chứng kiểm tra tín hiệu nền. Bên cạnh đó, xét nhu cầu lâm sàng, việc bổ sung hai loại mẫu dò đề dò hai vi khuẩn Campylobacter jejuni và Yersinia enterocolitica O:8 gây bệnh đường ruột chủ yếu ở trẻ em, là cần thiết. Quy trình PCR-RDB được hoàn thiện trong nghiên cứu này vì vậy có khả năng phát hiện đồng thời 14 vi khuẩn gây bệnh đường ruột, một cách đặc hiệu, với độ nhạy đạt 10^2 bản sao/ml, đã được thử nghiệm trên 30 mẫu phân, ghi nhận kết quả hoàn toàn khớp với kít thương mại PowerCheckTM 20 Pathogen Multiplex Real-time PCR Kit (Korea).


1996 ◽  
Vol 59 (8) ◽  
pp. 803-807 ◽  
Author(s):  
ROBERT G. REINHARD ◽  
T. JAMES McADAM ◽  
GEORGE J. FLICK ◽  
ROBERT E. CROONENBERGHS ◽  
ROBERT F. WITTMAN ◽  
...  

This study was conducted to determine the levels of Campylobacter jejuni, Campylobacter coli, Salmonella, Klebsiella pneumoniae, and Escherichia coli O157:H7 in fresh hand-picked blue crab (Callinectes sapidus) meat. An attempt was made to correlate these selected pathogens and general microbial quality to processors' sanitation practices and facility size. Hand-picked crabmeat samples from 12 blue-crab-processing facilities in the Chesapeake Bay region were collected and analyzed. Twenty samples from each of the different facilities were collected on different processing days. Facilities were chosen on the basis of production levels and Virginia Department of Health inspection scores as an indicator of sanitation practices (excellent, ≥94.5, acceptable, <94.5). All samples were tested between 16 h and 36 h after collection. Campylobacter jejuni was isolated from 36 (15%) of the 240 samples and Campylobacter coli was isolated from 14 (5.8%). Quantitative levels in all case were below limits of detection (<0.30 MPN/g). Klebsiella pneumoniae was isolated from a total of 51 (21%) samples. Counts ranged from less than 0.30 to 4.3 MPN/g. Aerobic plate counts ranged from 7.4 × 103 to 4.6 × 108 CFU/g with coliform counts ranging from <0.3 to 32.8 MPN/g. Fecal coliform levels were <0.3 to 2.26 MPN/g and Escherichia coli from <0.3 to 0.77 MPN/g. Salmonella spp. and Escherichia coli O157:H7 were not detected in any of the 240 samples analyzed. No significant differences (P < 0.05) between size and inspection scores were observed for general microbial quality or the presence of Klebsiella pneumoniae or Campylobacter species.


2000 ◽  
Vol 66 (12) ◽  
pp. 5472-5476 ◽  
Author(s):  
Vijay K. Sharma ◽  
Steve A. Carlson

ABSTRACT A multiplex fluorogenic PCR assay for simultaneous detection of pathogenic Salmonella strains and Escherichia coli O157:H7 was developed and evaluated for use in detecting very low levels of these pathogens in meat and feces. Two sets of primers were used to amplify a junctional segment of virulence genessipB and sipC of Salmonella and an intragenic segment of gene eae of E. coliO157:H7. Fluorogenic reporter probes were included in the PCR assay for automated and specific detection of amplified products. The assay could detect <10 CFU of Salmonella enterica serovar Typhimurium or E. coli O157:H7 per g of meat or feces artificially inoculated with these pathogens and cultured for 6 to 18 h in a single enrichment broth. Detection of amplification products could be completed in ≤4 h after enrichment.


2021 ◽  
pp. 103830
Author(s):  
Arícia Possas ◽  
Guiomar Denisse Posada-Izquierdo ◽  
Gonzalo Zurera ◽  
Fernando Pérez-Rodríguez

2019 ◽  
Vol 84 ◽  
pp. 103217 ◽  
Author(s):  
Susana de Oliveira Elias ◽  
Tiago Baptista Noronha ◽  
Eduardo Cesar Tondo

1999 ◽  
Vol 62 (4) ◽  
pp. 325-328 ◽  
Author(s):  
C. LITTLE ◽  
D. ROBERTS ◽  
E. YOUNGS ◽  
J. de LOUVOIS

A study of imported unprepared whole lettuces sampled from supermarkets, greengrocers, shops, and market stalls found that all were of acceptable microbiological quality. Twenty-seven out of 151 (18%) imported lettuce samples had Enterobacteriaceae levels of 104 CFU/g or more. However, these bacteria that constitute part of the natural microflora of unprepared vegetables may also be derived from the soil and/or by poor handling. The pathogens, Salmonella spp., Shigella spp., Campylobacter spp., Escherichia coli O157:H7, Vibrio cholerae, Listeria monocytogenes, and also Escherichia coli, an indicator of fecal contamination, were not detected in any imported lettuces, indicating that hygiene, harvesting, and production practices were good. Imported lettuces with Enterobacteriaceae levels of 104 CFU/g or more varied with type of retail premises and the temperature at which the lettuces were displayed. Samples from greengrocers, shops, and market stalls were more likely to contain Enterobacteriaceae at levels in excess of 104 CFU/g than those from supermarkets.


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