scholarly journals Rapid detection of Vibrio species using liquid microsphere arrays and real-time PCR targeting the ftsZ locus

2007 ◽  
Vol 56 (1) ◽  
pp. 56-65 ◽  
Author(s):  
Dobryan M. Tracz ◽  
Paul G. Backhouse ◽  
Adam B. Olson ◽  
Joanne K. McCrea ◽  
Julie A. Walsh ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Vibrio Parahaemolyticus ◽  
Rapid Detection ◽  
Vibrio Vulnificus ◽  
Comparative Sequence Analysis ◽  
Molecular Techniques ◽  
Array Technology ◽  
Species Specific ◽  
Pcr Targeting

The development of rapid and sensitive molecular techniques for the detection of Vibrio species would be useful for the surveillance of sporadic infections and management of major outbreaks. Comparative sequence analysis of the ftsZ gene in the predominant Vibrio species that cause human disease revealed distinct alleles for each examined species, including Vibrio cholerae, Vibrio parahaemolyticus and Vibrio vulnificus. Light Upon eXtension (LUX) real-time PCR assays were developed to target these species-specific polymorphisms, and were successful in rapidly differentiating the major pathogenic Vibrio species. Luminex liquid microsphere array technology was used to develop a comprehensive assay capable of simultaneously detecting V. cholerae, V. parahaemolyticus and V. vulnificus. These assays permitted the identification of a presumptive V. parahaemolyticus isolate as Vibrio alginolyticus, which was verified using additional molecular characterization.

scholarly journals Detection of Peanut Allergen Traces with a Real Time PCR Assay - The Challenge to Protect Food-Allergic Consumers

2017 ◽  
Vol 7 (1) ◽  
pp. 32 ◽  
Author(s):  
Dimitra Houhoula ◽  
Stamatios Koussissis ◽  
Vladimiros Lougovois ◽  
John Tsaknis ◽  
Dimitra Kassavita ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Rapid Detection ◽  
Food Products ◽  
Molecular Techniques ◽  
Food Allergens ◽  
Pcr Assay ◽  
Peanut Allergen ◽  
Pcr Method ◽  
Detection And Quantification

The aim of the present study was the implementation of molecular techniques in the detection and quantification of allergic substances of peanut in various kinds of food products, e.g., breakfast cereals, chocolates and biscuits that are frequently related to allergies. In some cases, the presence of peanuts can be due to contamination during production and are not declared on the label. A total of 152 samples were collected from supermarkets and were analysed by a Real Time PCR method. The results indicated that 125 samples (83,3%) were found positive in peanut traces but the most important finding is that from the 84 samples that had no allergen declaration for peanuts, 48 (57,1%) of them were found positive. In conclusion, Real Time PCR can be a very important tool for the rapid detection and quantification of food allergens.

Detection of enteropathogenic Vibrio parahaemolyticus, Vibrio cholerae and Vibrio vulnificus: performance of real-time PCR kits in an interlaboratory study

2017 ◽  
Vol 243 (8) ◽  
pp. 1335-1342 ◽  
Author(s):  
Erik Eschbach ◽  
Annett Martin ◽  
Jennifer Huhn ◽  
Constanze Seidel ◽  
Ralf Heuer ◽  
...  
Keyword(s):  
Vibrio Cholerae ◽  
Real Time ◽  
Real Time Pcr ◽  
Vibrio Parahaemolyticus ◽  
Vibrio Vulnificus ◽  
Interlaboratory Study

SYBR Green I-based real-time PCR targeting the rpoX gene for sensitive and rapid detection of Vibrio alginolyticus

2011 ◽  
Vol 25 (2-3) ◽  
pp. 137-141 ◽  
Author(s):  
Zhao Jing-jing ◽  
Chen Chang ◽  
Luo Peng ◽  
Ren Chun-hua ◽  
Jiang Xiao ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Rapid Detection ◽  
Vibrio Alginolyticus ◽  
Sybr Green ◽  
Sybr Green I ◽  
Pcr Targeting

Design and Validation of a Novel Multiplex Real-Time PCR Assay for Vibrio Pathogen Detection

2011 ◽  
Vol 74 (6) ◽  
pp. 939-948 ◽  
Author(s):  
ROBERT S. TEBBS ◽  
PIUS M. BRZOSKA ◽  
MANOHAR R. FURTADO ◽  
OLGA V. PETRAUSKENE
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Pathogen Detection ◽  
Nearest Neighbor ◽  
Vibrio Vulnificus ◽  
Molecular Techniques ◽  
Pcr Assay ◽  
Accurate Identification ◽  
Vibrio Pathogens ◽  
Pathogenic Vibrios

Three species—Vibrio cholerae, Vibrio parahaemolyticus, and Vibrio vulnificus—account for the majority of vibrio infections in humans. Rapid and accurate identification of Vibrio species has been problematic because phenotypic characteristics are variable within species. Additionally, biochemical identification and confirmation require 2 or more days to complete. Rapid and sensitive molecular techniques for the detection of vibrio pathogens would be useful for the surveillance and management of outbreaks. To facilitate the identification of human-pathogenic species, we designed and validated a highly sensitive, specific, and robust multiplex real-time PCR assay to identify V. cholerae, V. parahaemolyticus, and V. vulnificus using a four-dye configuration in a convenient lyophilized format. Multiple Vibrio strains were sequenced to verify candidate target TaqMan sites. Several individual assays within the multiplex contain multiple primers or probes to ensure detection of polymorphic variants. V. cholerae, V. parahaemolyticus, and V. vulnificus were detected either individually or in mixtures at ≤30 genomic copies. V. cholerae was specifically detected in the presence or absence of Vibrio mimicus. The Vibrio multiplex assay showed 100% specificity to all targets analyzed and no detection of nearest neighbor strains. Each assay exhibited 100% ± 10% efficiency. Multiplex real-time PCR can simplify pathogen detection and reduce costs per test since three species can be analyzed in a single reaction tube. Rapid and accurate detection of pathogenic vibrios in shellfish or seawater samples will improve the microbiological safety of seafood for consumers.

scholarly journals Serological and molecular evidence of Brucella species in the rapidly growing pig sector in Kenya

2020 ◽  
Author(s):  
James Miser Akoko ◽  
Roger Pelle ◽  
Velma Kivali ◽  
Esther Schelling ◽  
Gabriel Shirima ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Epidemiological Data ◽  
Molecular Techniques ◽  
Molecular Evidence ◽  
Rt Pcr ◽  
Growing Pig ◽  
Pcr Targeting ◽  
The Rose

Abstract BackgroundBrucellosis is an emerging, yet neglected zoonosis that has been reported in Kenya. Epidemiological data on brucellosis in ruminants is readily accessible; however, reports on brucellosis in pigs remain limited. This study sought to detect Brucella infection in pig serum by both serological and molecular techniques. Serum from 700 pigs randomly collected at a centralized abattoir in Nairobi, Kenya were screened in parallel, using the Rose Bengal Test (RBT) and competitive Enzyme-Linked Immuno-sorbent Assay (cELISA) for antibodies against Brucella spp.. All sera positive by RBT and 16 randomly selected negative samples were further tested using conventional PCR targeting bcsp31 gene and real-time PCR (RT-PCR) assays targeting IS711 and bcsp31 genes.ResultsA prevalence of 0.57% (n=4/700) was estimated using RBT. All RBT positive sera were also positive by both PCRs, while two sero-negative samples also tested positive on RT-PCR (n = 6/20). Brucella abortus was detected in four out of the six PCR positive samples through a multiple real-time PCR. ConclusionThe detection of antibodies against Brucella spp. and DNA in serum from slaughterhouse pigs indicate the presence of Brucella in pigs. Therefore, investigation of the epidemiology and role of pigs in the transmission of brucellosis in Kenya is needed. Further targeted studies would be useful to systematically quantify and identify the spp. of Brucella in pigs.

Molecular identification of Bartonella quintana infection using species-specific real-time PCR targeting transcriptional regulatory protein (bqtR) gene

2011 ◽  
Vol 25 (5-6) ◽  
pp. 238-242 ◽  
Author(s):  
Maria Carla Liberto ◽  
Angelo G. Lamberti ◽  
Nadia Marascio ◽  
Giovanni Matera ◽  
Angela Quirino ◽  
...  
Keyword(s):  
Real Time ◽  
Molecular Identification ◽  
Real Time Pcr ◽  
Regulatory Protein ◽  
Bartonella Quintana ◽  
Transcriptional Regulatory ◽  
Species Specific ◽  
Pcr Targeting

scholarly journals Considerations for incorporating real-time PCR assays into routine marine biosecurity surveillance programmes: a case study targeting the Mediterranean fanworm (Sabella spallanzanii) and club tunicate (Styela clava)

Genome ◽  
2019 ◽  
Vol 62 (3) ◽  
pp. 137-146 ◽  
Author(s):  
Susanna A. Wood ◽  
Xavier Pochon ◽  
Witold Ming ◽  
Ulla von Ammon ◽  
Chris Woods ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Molecular Techniques ◽  
Extracellular Dna ◽  
Occupancy Modelling ◽  
Styela Clava ◽  
Dna And Rna ◽  
Marine Invasive Species ◽  
Pcr Assays ◽  
Species Specific

Molecular techniques may provide effective tools to enhance marine biosecurity surveillance. Prior to routine implementation, evidence-based consideration of their benefits and limitations is needed. In this study, we assessed the efficiency and practicality of visual diver surveys and real-time PCR assays (targeting DNA and RNA) for detecting two marine invasive species whose infestation levels varied between species and location: Sabella spallanzanii and Styela clava. Filtered water samples (n = 171) were collected in parallel with dive surveys at two locations as part of the New Zealand Marine High Risk Site Surveillance programme: Nelson Harbour (27 sites) and Waitemata Harbour (30 sites). Diver surveys resulted in a greater number of detections compared to real-time PCR: S. clava – 21 versus 5 sites in Nelson, 6 versus 1 in Auckland; S. spallanzanii – 18 versus 10 in Auckland, no detections in Nelson. Occupancy modelling derived detection probabilities for the real-time PCR for S. clava were low (14%), compared to S. spallanzanii (66%). This could be related to abundances, or species-specific differences in DNA shedding. Only one RNA sample was positive, suggesting that most detections were from extracellular DNA or non-viable fragments. While molecular methods cannot yet replace visual observations, this study shows they provide useful complementary information.

Sign in / Sign up

Export Citation Format

Share Document