Simultaneous detection and differentiation of human parvovirus B19 and human parvovirus 4 by an internally controlled multiplex quantitative real-time PCR

2017 ◽  
Vol 36 ◽  
pp. 50-57
Author(s):  
Junting Jia ◽  
Yadi Zhong ◽  
Yi Guo ◽  
Chaoji Huangfu ◽  
Xiong Zhao ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Parvovirus B19 ◽  
Simultaneous Detection ◽  
Human Parvovirus B19 ◽  
Quantitative Real Time Pcr

Quantitative real-time PCR for differential diagnostics of parvovirus B19 infection in acute liver failure patients

2019 ◽  
Vol 19 (3) ◽  
pp. 259-266 ◽  
Author(s):  
Arthur Daniel Rocha Alves ◽  
Rita De Cassia Nasser Cubel Garcia ◽  
Oswaldo Gonçalves Cruz ◽  
Marcelo Alves Pinto ◽  
Luciane Almeida Amado Leon
Keyword(s):  
Liver Failure ◽  
Acute Liver Failure ◽  
Real Time ◽  
Real Time Pcr ◽  
Parvovirus B19 ◽  
Differential Diagnostics ◽  
Quantitative Real Time Pcr ◽  
Parvovirus B19 Infection

Simultaneous detection and quantification of 19 diarrhea-related pathogens with a quantitative real-time PCR panel assay

2018 ◽  
Vol 151 ◽  
pp. 76-82 ◽  
Author(s):  
Warawan Wongboot ◽  
Kazuhisa Okada ◽  
Siriporn Chantaroj ◽  
Watcharaporn Kamjumphol ◽  
Shigeyuki Hamada
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Simultaneous Detection ◽  
Quantitative Real Time Pcr ◽  
Detection And Quantification

scholarly journals Pentaplexed Quantitative Real-Time PCR Assay for the Simultaneous Detection and Quantification of Botulinum Neurotoxin-Producing Clostridia in Food and Clinical Samples

2010 ◽  
Vol 76 (13) ◽  
pp. 4387-4395 ◽  
Author(s):  
Sebastian Kirchner ◽  
K. Melanie Krämer ◽  
Martin Schulze ◽  
Diana Pauly ◽  
Daniela Jacob ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Target Genes ◽  
False Negative ◽  
Simultaneous Detection ◽  
Locked Nucleic Acid ◽  
Clinical Samples ◽  
Quantitative Real Time Pcr ◽  
Pcr Assay ◽  
Pcr Assays

ABSTRACT Botulinum neurotoxins are produced by the anaerobic bacterium Clostridium botulinum and are divided into seven distinct serotypes (A to G) known to cause botulism in animals and humans. In this study, a multiplexed quantitative real-time PCR assay for the simultaneous detection of the human pathogenic C. botulinum serotypes A, B, E, and F was developed. Based on the TaqMan chemistry, we used five individual primer-probe sets within one PCR, combining both minor groove binder- and locked nucleic acid-containing probes. Each hydrolysis probe was individually labeled with distinguishable fluorochromes, thus enabling discrimination between the serotypes A, B, E, and F. To avoid false-negative results, we designed an internal amplification control, which was simultaneously amplified with the four target genes, thus yielding a pentaplexed PCR approach with 95% detection probabilities between 7 and 287 genome equivalents per PCR. In addition, we developed six individual singleplex real-time PCR assays based on the TaqMan chemistry for the detection of the C. botulinum serotypes A, B, C, D, E, and F. Upon analysis of 42 C. botulinum and 57 non-C. botulinum strains, the singleplex and multiplex PCR assays showed an excellent specificity. Using spiked food samples we were able to detect between 103 and 105 CFU/ml, respectively. Furthermore, we were able to detect C. botulinum in samples from several cases of botulism in Germany. Overall, the pentaplexed assay showed high sensitivity and specificity and allowed for the simultaneous screening and differentiation of specimens for C. botulinum A, B, E, and F.

A Quantitative Real-Time PCR Approach for Assessing Campylobacter jejuni and Campylobacter coli Colonization in Broiler Herds

2017 ◽  
Vol 80 (4) ◽  
pp. 604-608 ◽  
Author(s):  
Katrin Haas ◽  
Gudrun Overesch ◽  
Peter Kuhnert
Keyword(s):  
Real Time ◽  
Campylobacter Jejuni ◽  
Real Time Pcr ◽  
Simultaneous Detection ◽  
Developed Countries ◽  
Human Infection ◽  
Health Concern ◽  
Campylobacter Coli ◽  
Quantitative Real Time Pcr ◽  
Simple Method

ABSTRACT Human campylobacteriosis is a major public health concern in developed countries, with Campylobacter jejuni and Campylobacter coli from poultry recognized as the main source of human infection. Identification of Campylobacter-positive broiler herds before slaughter is essential for implementing measures to avoid carryover of pathogens via the slaughter process into the food chain. However, appropriate methods that have been validated for testing poultry flocks antemortem are lacking for Campylobacter. A quantitative real-time PCR (qPCR) that allows simultaneous detection and quantification of C. jejuni and C. coli was adapted and optimized to be applied on boot socks. The adjusted qPCR serves as an easy, sensitive, and quantitative method for Campylobacter detection in poultry flocks antemortem by analysis of boot socks. An adequate correlation was found between qPCR and culture, as well as between boot socks and cecal samples, which are regarded as the “gold standard.” Therefore, boot sock sampling followed by qPCR analysis provides a reliable and simple method for assessing Campylobacter load within a flock prior to slaughter. The approach allows categorization of broiler herds into negative, low, moderate, or high Campylobacter colonization. Based on the results of this new approach, risk assessment models, such as evaluating the possible effect of sorting flocks before slaughter, can be easily implemented. Similarly, targeted identification of highly colonized flocks for improvement of biosecurity measures at the farm level will become feasible, presenting an opportunity to increase food safety.

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