scholarly journals Development of Rapid Screening of Campylobacter spp. and the GyrA Mutants Using PCR-DNA Strip and Real-Time PCR

2016 ◽  
Vol 33 (1) ◽  
pp. 12-18
Author(s):  
Chiaki KOJIMA ◽  
Michiru KISHIMOTO ◽  
Machiko MIYATA ◽  
Sayumi NOMURA ◽  
Takayuki EZAKI
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Rapid Screening

scholarly journals A Real-Time PCR Screening Assay for Rapid Detection of Listeria Monocytogenes Outbreak Strains

Foods ◽  
2020 ◽  
Vol 9 (1) ◽  
pp. 67 ◽  
Author(s):  
Marina Torresi ◽  
Anna Ruolo ◽  
Vicdalia Aniela Acciari ◽  
Massimo Ancora ◽  
Giuliana Blasi ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Real Time ◽  
Real Time Pcr ◽  
Human Case ◽  
Outbreak Investigation ◽  
Rapid Screening ◽  
Reference Laboratory ◽  
Screening Assay ◽  
Pcr Screening ◽  
Umbria Region

From January 2015 to March 2016, an outbreak of 23 human cases of listeriosis in the Marche region and one human case in the Umbria region of Italy was caused by Listeria monocytogenes strains showing a new pulsotype never described before in Italy. A total of 37 clinical strains isolated from patients exhibiting listeriosis symptoms and 1374 strains correlated to the outbreak were received by the Italian National Reference Laboratory for L. monocytogenes (It NRL Lm) of Istituto Zooprofilattico Sperimentale dell’Abruzzo e del Molise (IZSAM) for outbreak investigation. A real-time PCR assay was purposely designed for a rapid screening of the strains related to the outbreak. PCR-positive strains were successively typed through molecular serogrouping, pulsed field gel electrophoresis (PFGE), and Next Generation Sequencing (NGS). Applying the described strategy, based on real-time PCR screening, we were able to considerably reduce time and costs during the outbreak investigation activities.

Rapid Screening of Leukemia Fusion Transcripts in Acute Leukemia by Real-time PCR

2002 ◽  
Vol 43 (12) ◽  
pp. 2291-2299 ◽  
Author(s):  
Kazuoki Osumi ◽  
Takafumi Fukui ◽  
Hitoshi Kiyoi ◽  
Masanobu Kasai ◽  
Yoshihisa Kodera ◽  
...  
Keyword(s):  
Acute Leukemia ◽  
Real Time ◽  
Real Time Pcr ◽  
Rapid Screening ◽  
Fusion Transcripts

Improved diagnostic and real-time pcr in rapid screening for Salmonella in the poultry food chain

2006 ◽  
Vol 54 (3) ◽  
pp. 297-312 ◽  
Author(s):  
Annamária Szmolka ◽  
Éva Kaszanyitzky ◽  
B. Nagy
Keyword(s):  
Real Time ◽  
Food Chain ◽  
Real Time Pcr ◽  
Culture Method ◽  
Rapid Screening ◽  
Poultry Feed ◽  
Broiler Chicks ◽  
Diagnostic Pcr ◽  
Enrichment Cultures ◽  
Specific Pcr

The goal of this study was to improve the diagnostic applicability of genus- and serovar- (S. Enteritidis and S. Typhimurium) specific PCR systems in screening faecal and caecal samples of poultry, poultry feed and poultrymeat for Salmonella, by keeping the opportunity to obtain Salmonella cultures from positive samples. Peptone broth pre-enrichment cultures of the samples were tested by PCR. In faecal and caecal samples from broiler chicks a strong inhibitory action was frequently observed. This could be reduced markedly by the addition of bovine serum albumin (BSA) acting as amplification facilitator. The results of testing pre-enrichment cultures from artificially contaminated faecal, poultry feed and poultrymeat samples (using S. Enteritidis, S. Typhimurium and S. Hadar as contaminants) suggest that the sensitivity of the above systems is 101-102 CFU g-1 sample. The testing of 95 caecal samples from slaughtered chicks resulted in 49% culture-positive and 76% PCR-positive samples. The suitability of a generic real-time PCR for testing faecal samples of poultry was also studied. Its detection limit for these samples was found to be lower than that of the diagnostic PCR system. Both methods reduced the time required for Salmonella detection to 24-30 h, and the advantage of the real-time PCR was its increased sensitivity. We have established a diagnostic and a real-time PCR system for rapid and reliable genus- and serovar- (S. Enteritidis and S. Typhimurium) specific detection of Salmonella for monitoring purposes in the poultry food chain. Sensitivity is equal to, or higher than, that of the standard bacterial culture method, and the method still provides the Salmonella culture if needed.

scholarly journals Rapid Screening Method for Compounds That Affect the Growth and Germination of Candida albicans, Using a Real-Time PCR Thermocycler

2011 ◽  
Vol 77 (22) ◽  
pp. 8193-8196 ◽  
Author(s):  
Lucja M. Jarosz ◽  
Bastiaan P. Krom
Keyword(s):  
Candida Albicans ◽  
Real Time ◽  
Real Time Pcr ◽  
Fluorescent Protein ◽  
Screening Method ◽  
Rapid Screening ◽  
Content Type ◽  
Wide Range ◽  
Hyphal Formation ◽  
Green Fluorescent

ABSTRACTWe propose a screening method for compounds affecting growth and germination inCandida albicansusing a real-time PCR thermocycler to quantify green fluorescent protein (GFP) fluorescence. Using PACT1-GFPand PHWP1-GFPreporter strains, the effects of a wide range of compounds on growth and hyphal formation were quantitatively assessed within 3 h after inoculation.

Rapid screening of waterborne pathogens using phage-mediated separation coupled with real-time PCR detection

2016 ◽  
Vol 408 (15) ◽  
pp. 4169-4178 ◽  
Author(s):  
Ziyuan Wang ◽  
Danhui Wang ◽  
Amanda J. Kinchla ◽  
David A. Sela ◽  
Sam R. Nugen
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Pcr Detection ◽  
Rapid Screening ◽  
Waterborne Pathogens

Direct urine polymerase chain reaction for chlamydia and gonorrhoea: a simple means of bringing high-throughput rapid testing to remote settings?

Sexual Health ◽  
2013 ◽  
Vol 10 (4) ◽  
pp. 299 ◽  
Author(s):  
Frashta Rahimi ◽  
Namraj Goire ◽  
Rebecca Guy ◽  
John M. Kaldor ◽  
James Ward ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
High Throughput ◽  
Real Time Pcr ◽  
Point Of Care ◽  
Resistance Mechanisms ◽  
Rapid Screening ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Urine Testing

Background Rapid point-of-care tests (POCTs) for chlamydia (Chlamydia trachomatis) and gonorrhoea (Neisseria gonorrhoeae) have the potential to confer health benefits in certain populations even at moderate sensitivities; however, suitable POCTs for these organisms are currently lacking. Methods: In this study, we investigated the use of direct urine polymerase chain reaction (PCR), with the view of implementing a simplified PCR strategy for high-throughput chlamydia and gonorrhoea screening in remote settings. Briefly, a simple dilution of the urine was performed before adding it directly to a real-time PCR reaction. The method was evaluated using 134 stored urine specimens that had been submitted for chlamydia and gonorrhoea testing and had been tested using a commercial C. trachomatis and N. gonorrhoeae PCR method. These included samples that were PCR-positive for chlamydia (n = 87), gonorrhoea (n = 16) or both (n = 2). Direct urine testing was conducted using previously described in-house real-time PCR methods for C. trachomatis and N. gonorrhoeae as well as for recognised N.gonorrhoeae antimicrobial resistance mechanisms. Results: The overall sensitivities and specificities of the direct urine PCR were 78% and 100% for chlamydia, and 83% and 100% for gonorrhoea. N.gonorrhoeae penicillin and quinolone resistance mechanisms were characterised in 14 of the 18 N. gonorrhoeae-positive samples. Conclusions: The results of this study show that the simplified PCR strategy may be a feasible approach for rapid screening and improving chlamydia and gonorrhoea treatment in remote settings.

scholarly journals Laboratory Analysis of an Outbreak of Candida auris in New York from 2016 to 2018-Impact and Lessons Learned

2019 ◽  
Author(s):  
YanChun Zhu ◽  
Brittany O’Brien ◽  
Lynn Leach ◽  
Alexandra Clark ◽  
Marian Bates ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Antifungal Susceptibility ◽  
Clinical Isolates ◽  
Rapid Screening ◽  
Care Hospital ◽  
Environmental Surveillance ◽  
Intrinsic Resistance ◽  
Candida Auris ◽  
Healthcare Facilities

ABSTRACTCandida auris is a multidrug-resistant yeast which has emerged in healthcare facilities worldwide, however little is known about identification methods, patient colonization, spread, environmental survival, and drug resistance. Colonization on both biotic and abiotic surfaces, along with travel, appear to be the major factors for the spread of this pathogen across the globe. In this investigation, we present laboratory findings from an ongoing C. auris outbreak in NY from August 2016 through 2018. A total of 540 clinical isolates, 11,035 patient surveillance specimens, and 3,672 environmental surveillance samples were analyzed. Laboratory methods included matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) for yeast isolate identification, real-time PCR for rapid surveillance sample screening, culture on selective/non-selective media for recovery of C. auris and other yeasts from surveillance samples, antifungal susceptibility testing to determine the C. auris resistance profile, and Sanger sequencing of ribosomal genes for C. auris genotyping. Results included: a) identification and confirmation of C. auris in 413 clinical isolates and 931 patient surveillance isolates, as well as identification of 277 clinical cases and 350 colonized cases from 151 healthcare facilities including 59 hospitals, 92 nursing homes, 1 long-term acute care hospital (LTACH), and 2 hospices, b) successful utilization of an in-house developed C. auris real-time PCR assay for the rapid screening of patient and environmental surveillance samples, c) demonstration of relatively heavier colonization of C. auris in nares compared to the axilla/groin, and d) predominance of the South Asia Clade I with intrinsic resistance to fluconazole and elevated minimum inhibitory concentration (MIC) to voriconazole (81%), amphotericin B (61%), 5-FC (3%) and echinocandins (1%). These findings reflect greater regional prevalence and incidence of C. auris and the deployment of better detection tools in an unprecedented outbreak.

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