Real-time PCR-based microfluidic array chip for simultaneous detection of multiple waterborne pathogens

Ramalingam, Naveen; Zhang, Rui; Liu, Hao-Bing; Dai, Changchun; Kaushik, Rajni; Ratnaharika, Bandi; Gong, Haiqing

doi:10.1016/j.snb.2009.11.025

Cited by 66 publications

(36 citation statements)

References 22 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In addition, microfluidic technology has also been used for the simultaneous detection of multiple pathogens (10,44). For example, Stedtfeld et al (44) used the Biotrove OpenArray system (Applied Biosystems) to run SYBR green qPCR assays to detect 30 pathogens.…”

Section: Discussionmentioning

confidence: 99%

“…Although these approaches greatly reduce the detection time compared with conventional culture-based methods, the majority of them do not provide quantitative information on the target pathogens present in samples. Some quantitative (or semiquantitative) methods are available (10,11,13,(16)(17)(18)20), but they are not sensitive enough to detect small amounts of pathogens. In addition, their specificity is also limited, especially in 16S rRNA gene-based microarrays and pyrosequencing, as some pathogens are indistinguishable from nonpathogens based on their 16S rRNA gene sequence information alone (e.g., general E. coli versus enterohemorrhagic E. coli [EHEC]).…”

mentioning

confidence: 99%

See 1 more Smart Citation

Simultaneous Quantification of Multiple Food- and Waterborne Pathogens by Use of Microfluidic Quantitative PCR

Ishii

Segawa

Okabe

2013

Appl Environ Microbiol

122

159

View full text Add to dashboard Cite

bThe 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 (general Escherichia 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.

show abstract

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Simultaneous Quantification of Multiple Food- and Waterborne Pathogens by Use of Microfluidic Quantitative PCR

Ishii

Segawa

Okabe

2013

Appl Environ Microbiol

122

159

View full text Add to dashboard Cite

show abstract

“…101 In 2010, Ramalingam and colleagues presented the simultaneous detection of four waterborne bacteria in a PDMS PCR array, which utilized capillary flow for sample loading. 102 In 2012, Agrawal and coworkers showed simultaneous detection of E. coli and S. tyhimurium on a circular microfluidics design manufactured in PDMS using magnetic nanoparticles to control and concentrate samples. 61 With regard to protozoa, two publications that relate to molecular sensing of Cryptosporidium in miniaturized format describe the performance of NASBA off-chip 103 ; only the detection of the mRNAs amplicons was performed on-chip.…”

Section: Waterborne Pathogensmentioning

confidence: 99%

Miniaturized Detection Systems

Bridle¹

2014

Waterborne Pathogens

View full text Add to dashboard Cite

“…A disposable microfluidic chip with 9 reaction channels made from polydimethylsiloxane and glass was developed for parallel detection of waterborne pathogens (Ramalingam et al 2010 (Seyrig et al 2010).…”

Section: Polymerase Chain Reaction Assays Formentioning

confidence: 99%

Detection and Occurrence of Indicator Organisms and Pathogens

Seyrig¹,

Herzog²,

Ahmad³

et al. 2011

Water Environment Research

View full text Add to dashboard Cite

This review summarizes the studies related to detection and occurrence of fecal indicator bacteria (FIB) and pathogens published during 2010. The first section of this review summarizes various detection methods for waterborne pathogens including endpoint, reverse transcriptase, and real-time polymerase chain reaction (PCR), loop-mediated isothermal amplification (LAMP), microarrays, and immunoassays. Relevant literature on sample processing is also included. The second section focuses on phenotypic and genotypic methods for microbial source tracking. The third section summarizes studies related to occurrence, persistence, and transport with emphasis on microbial quality of recreational beaches and watersheds.

show abstract

Real-time PCR-based microfluidic array chip for simultaneous detection of multiple waterborne pathogens

Cited by 66 publications

References 22 publications

Simultaneous Quantification of Multiple Food- and Waterborne Pathogens by Use of Microfluidic Quantitative PCR

Simultaneous Quantification of Multiple Food- and Waterborne Pathogens by Use of Microfluidic Quantitative PCR

Miniaturized Detection Systems

Detection and Occurrence of Indicator Organisms and Pathogens

Contact Info

Product

Resources

About