Segmented continuous-flow multiplex polymerase chain reaction microfluidics for high-throughput and rapid foodborne pathogen detection

2014 ◽  
Vol 826 ◽  
pp. 51-60 ◽  
Author(s):  
Bowen Shu ◽  
Chunsun Zhang ◽  
Da Xing
Keyword(s):  
Polymerase Chain Reaction ◽  
High Throughput ◽  
Continuous Flow ◽  
Multiplex Polymerase Chain Reaction ◽  
Pathogen Detection ◽  
Foodborne Pathogen ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Foodborne Pathogen Detection

scholarly journals Impact of Multiplex Polymerase Chain Reaction Testing for Respiratory Pathogen Detection in Pediatric Patients

2017 ◽  
Vol 4 (suppl_1) ◽  
pp. S503-S503
Author(s):  
Courtney C Sutton ◽  
Patti J Walton ◽  
Montgomery F Williams ◽  
Tracey L Bastian ◽  
Michael Wright ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Multiplex Polymerase Chain Reaction ◽  
Pathogen Detection ◽  
Pediatric Patients ◽  
Respiratory Pathogen ◽  
Chain Reaction ◽  
Polymerase Chain Reaction Testing ◽  
Polymerase Chain

scholarly journals Evaluation of a multiplex polymerase chain reaction for early diagnosis of ventriculostomy-related infections

2015 ◽  
Vol 123 (6) ◽  
pp. 1586-1592 ◽  
Author(s):  
Claire L. Gordon ◽  
Rafal Tokarz ◽  
Thomas Briese ◽  
W. Ian Lipkin ◽  
Komal Jain ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Multiplex Polymerase Chain Reaction ◽  
Pathogen Detection ◽  
High Sensitivity ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Clinical Value ◽  
Parallel Testing ◽  
Diagnostic Approaches ◽  
Polymerase Chain

OBJECT Diagnosis of ventriculostomy-related infections (VRIs) is challenging due to the lack of rapid, sensitive assays for pathogen detection. The authors report the development of a multiplex polymerase chain reaction (PCR) assay for differential diagnosis of common VRI pathogens. METHODS MassTag PCR was used to develop a multiplex assay for detection of 11 VRI pathogens. The assay was established and optimized using cloned template standards and spiked samples and was then evaluated on CSF specimens from ventricular drains. Subjects were grouped into definite VRI, possible VRI, or no VRI based on conventional microbiology, CSF evaluation, and clinical parameters. RESULTS CSF specimens were obtained from 45 subjects (median age 49 years, interquartile range 32–63 years; 51% were male). The assay detected 10–100 genome copies. It detected a pathogen in 100% (6 of 6) of definite VRI cases in which a pathogen targeted by the assay was present; these represented 67% of all definite VRIs (6 of 9). Among subjects with a possible VRI, the assay detected a pathogen in 29% (5 of 17). In subjects without overt infection the presence of a pathogen was detected in 32% of subjects (6 of 19), albeit with lower signal compared with the VRI group. CONCLUSIONS MassTag PCR enabled parallel testing of CSF specimens for 11 pathogens of VRI. The high sensitivity of PCR combined with possible device colonization, specimen contamination, and concurrent antibiotic treatments limit the clinical value of the assay, similar to other current diagnostic approaches. With further optimization, multiplex PCR may provide timely identification of multiple possible VRI pathogens and guide management, complementing classic culture approaches.

scholarly journals Multiplex polymerase chain reaction pathogen detection in patients with suspected septicemia after trauma, emergency, and burn surgery

Surgery ◽  
2012 ◽  
Vol 151 (3) ◽  
pp. 456-463 ◽  
Author(s):  
Nam K. Tran ◽  
David H. Wisner ◽  
Timothy E. Albertson ◽  
Stuart Cohen ◽  
David Greenhalgh ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Multiplex Polymerase Chain Reaction ◽  
Pathogen Detection ◽  
Chain Reaction ◽  
Burn Surgery ◽  
Polymerase Chain

scholarly journals A Novel Capillary Electrophoresis-Based High-Throughput Multiplex Polymerase Chain Reaction System for the Simultaneous Detection of Nine Pathogens in Swine

2017 ◽  
Vol 2017 ◽  
pp. 1-8 ◽  
Author(s):  
Xu-long Wu ◽  
Lu Xiao ◽  
Hua Lin ◽  
Miao Yang ◽  
Shi-jie Chen ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Capillary Electrophoresis ◽  
Detection Limit ◽  
High Throughput ◽  
Multiplex Polymerase Chain Reaction ◽  
Simultaneous Detection ◽  
Reaction System ◽  
Clinical Samples ◽  
Chain Reaction ◽  
Polymerase Chain

Here we aimed to develop a capillary electrophoresis-based high-throughput multiplex polymerase chain reaction (PCR) system for the simultaneous detection of nine pathogens in swine. Nine pairs of specific primers and a set of universal primers were designed; the multiplex PCR was established. The specificity and cross-reactivity of this assay were examined, and the detection limit was determined using serial 10-fold dilutions of plasmids containing the target sequences. The assay was further tested using 144 clinical samples. We found that the nine specific amplification peaks were observed, and the assay had a high degree of specificity, without nonspecific amplification. The simultaneous detection limit for the nine viruses reached 10000 copies μL−1 when all of the premixed viral targets were present. Seventy-seven of the clinical samples tested positive for at least one of the viruses; the principal viral infections in the clinical samples were porcine circovirus type 2 and porcine reproductive and respiratory syndrome virus. This approach has much potential for further development of high-throughput detection tools for the diagnosis of diseases in animals.

A handheld continuous-flow real-time fluorescence qPCR system with a PVC microreactor

The Analyst ◽  
2020 ◽  
Vol 145 (7) ◽  
pp. 2767-2773 ◽  
Author(s):  
Bing Shi ◽  
Yuanming Li ◽  
Di Wu ◽  
Wenming Wu
Keyword(s):  
Polymerase Chain Reaction ◽  
Molecular Biology ◽  
Real Time ◽  
Continuous Flow ◽  
Pathogen Detection ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Sensitivity Specificity

The polymerase chain reaction (PCR) has unique advantages of sensitivity, specificity and rapidity in pathogen detection, which makes it at the forefront of academia and application in molecular biology diagnosis.

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