A multiplex nanoparticle-based bio-barcoded DNA sensor for the simultaneous detection of multiple pathogens

2010 ◽  
Vol 26 (4) ◽  
pp. 1736-1742 ◽  
Author(s):  
Deng Zhang ◽  
Michael C. Huarng ◽  
Evangelyn C. Alocilja
2018 ◽  
Vol 265 ◽  
pp. 514-521 ◽  
Author(s):  
Sakda Jampasa ◽  
Weena Siangproh ◽  
Rawiwan Laocharoensuk ◽  
Pattamawadee Yanatatsaneejit ◽  
Tirayut Vilaivan ◽  
...  

2019 ◽  
Vol 14 (10) ◽  
pp. 885-898 ◽  
Author(s):  
Moezi Parichehr ◽  
Kargar Mohammad ◽  
Doosti Abbas ◽  
Khoshneviszadeh Mehdi

Aim: The aim of this study is to formulate a new single nonselective pre-enrichment medium (ELSS) that can support the concurrent growth of four major foodborne pathogens containing E. coli O157: H7, L. monocytogenes, S. aureus and S. enterica serovar Entertidis to develop a multiplex TaqMan Real-time PCR (mRT-PCR). Methods: The mRT-PCR with a new pre-enrichment was carried out for simultaneous detection and quantification of these foodborne bacteria. Results: By using mRT-PCR after 16 h pre-enrichment in ELSS, the detection limit of each pathogen was 1 CFU/25 ml contaminated milk, as well as inclusivity and exclusivity reached 100%. Conclusion: The mRT-PCR assay with pre-enrichment step is a fast and reliable technique for detecting single or multiple pathogens in food products.


2017 ◽  
Vol 5 (1) ◽  
pp. 57-66 ◽  
Author(s):  
Zeeshan Ali ◽  
Jiuhai Wang ◽  
Yongjun Tang ◽  
Bin Liu ◽  
Nongyue He ◽  
...  

In this report, a DNA hybridization based chemiluminescent detection method has been proposed for reliable detection of multiple pathogens. The use of surface modified magnetic nanoparticles can help to integrate this system into an automated platform for high throughput applications.


2017 ◽  
Vol 52 (2) ◽  
pp. 186-195 ◽  
Author(s):  
Hsiang-Yun Tung ◽  
Wei-Chen Chen ◽  
Bor-Rung Ou ◽  
Jan-Ying Yeh ◽  
Yeong-Hsiang Cheng ◽  
...  

Traditional serological enzyme-linked immunosorbent assay (ELISA) is routinely used to monitor pathogens during quarantine in most animal facilities to prevent possible infection. However, the ELISA platform is a single-target assay, and screening all targeted pathogens is time-consuming and laborious. In this study, to increase sensitivity and to reduce diagnosis time for high-throughput processes, multiplex PCR and DNA biochip techniques were combined to develop a multi-pathogen diagnostic method for use instead of routine ELISA. Eight primer sets were designed for multiplex PCR to detect genes from seven targeted bacterial and viral pathogens. DNA–DNA hybridization was conducted on a biochip following the multiple PCR analysis. Using this method, a total of 24 clinical samples were tested, and the result showed that not only single infection but also co-infection by multi-pathogens can be detected. In conclusion, multiplex PCR coupled with a DNA biochip is an efficient method for detecting multi-pathogens in a reaction. This platform is a useful tool for quarantine services and disease prevention in animal facilities.


2018 ◽  
Vol 90 (15) ◽  
pp. 9621-9628 ◽  
Author(s):  
Linyao Li ◽  
Qingjin Li ◽  
Ziyi Liao ◽  
Yan Sun ◽  
Quansheng Cheng ◽  
...  

2017 ◽  
Vol 4 (suppl_1) ◽  
pp. S17-S18
Author(s):  
Jennifer Milucky ◽  

Abstract Background Pneumonia causes significant morbidity and mortality worldwide. Comprehensive etiology studies of pneumonia in adults are limited; however, new diagnostics enable simultaneous detection of multiple pathogens in respiratory specimens. Characterizing the public health threat of severe acute respiratory infection (SARI) may enhance global health security. We studied potential etiologies of SARI among adults in six countries over a 12-month period using multi-pathogen diagnostics. Methods We enrolled SARI cases (acute onset of fever and cough, requiring hospitalization, in an adult) from Global Disease Detection sites in Bangladesh, China, Egypt, Guatemala, Kenya, and Thailand and healthy frequency-matched controls (2 controls: 5 cases) by time (onset), age group (18–49, 50–64, 65+ years), and catchment area. Demographics, clinical data, and nasopharyngeal and oropharyngeal specimens were collected from cases and controls. Specimens were tested for 16 viruses and 14 bacteria using Taqman® Array Card, which uses real-time reverse transcriptase polymerase chain reaction. Results We enrolled 2,388 cases and 1,135 controls from Oct 2013 through Oct 2015. Age distribution (Figure) and seasonality varied by site: enrollment peaked in summer months in Bangladesh, Thailand, and China, and in winter months in Egypt, but was stable throughout the year in Guatemala and Kenya. Case fatality rate across all study locations was 2.3% (range 0–7.0%). One or more pathogens was detected in 76% of cases and in 67% of controls; ≥2 pathogens were detected in 42% of cases and 37% of controls. Pathogens more commonly detected among cases than controls included influenza A (OR 13.3, CI 7.0–25.2; 12.8% of cases vs. 1.1% of controls), influenza B (OR: 27.0, CI 8.6–84.8; 8.1% vs. 0.3%), and respiratory syncytial virus (RSV) (OR: 9.4, CI 3.4–25.8; 4.0% vs. 0.4%). Conclusion In this SARI study, frequent detection of multiple pathogens in the oro- and nasopharynx of both cases and controls made etiology attribution difficult. Influenza and RSV, however, were likely to be causes of SARI. Because upper respiratory tract specimens may not accurately reflect disease in the lung, better specimens are needed to determine pneumonia etiology, particularly for bacteria. Disclosures All authors: No reported disclosures.


2013 ◽  
Vol 79 (9) ◽  
pp. 2891-2898 ◽  
Author(s):  
Satoshi Ishii ◽  
Takahiro Segawa ◽  
Satoshi Okabe

ABSTRACTThe direct quantification of multiple pathogens has been desired for diagnostic and public health purposes for a long time. In this study, we applied microfluidic quantitative PCR (qPCR) technology to the simultaneous detection and quantification of multiple food- and waterborne pathogens. In this system, multiple singleplex qPCR assays were run under identical detection conditions in nanoliter-volume chambers that are present in high densities on a chip. First, we developed 18 TaqMan qPCR assays that could be run in the same PCR conditions by using prevalidated TaqMan probes. Specific and sensitive quantification was achieved by using these qPCR assays. With the addition of two previously validated TaqMan qPCR assays, we used 20 qPCR assays targeting 10 enteric pathogens, a fecal indicator bacterium (generalEscherichia coli), and a process control strain in the microfluidic qPCR system. We preamplified the template DNA to increase the sensitivity of the qPCR assays. Our results suggested that preamplification was effective for quantifying small amounts of the template DNA without any major impact on the sensitivity, efficiency, and quantitative performance of qPCR. This microfluidic qPCR system allowed us to detect and quantify multiple pathogens from fecal samples and environmental water samples spiked with pathogens at levels as low as 100 cells/liter. These results suggest that the routine monitoring of multiple pathogens in food and water samples is now technically feasible. This method may provide more reliable information for risk assessment than the current fecal contamination indicator approach.


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