Automated methods for multiplexed pathogen detection

Straub, Timothy M.; Dockendorff, Brian P.; Quinonez-Diaz, Maria D.; Valdez, Catherine O.; Shutthanandan, Janani I.; Tarasevich, Barbara J.; Grate, Jay W.; Bruckner-Lea, Cynthia J.

doi:10.1016/j.mimet.2005.04.012

Cited by 64 publications

(38 citation statements)

References 40 publications

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“…This observation is corroborated by the more variable and less sensitive results obtained from non-enriched samples of water, soil, and tomato wash (Table 2). However, the standard culturebased technique could be essentially as sensitive as the enriched qPCR method, but three major disadvantages were identified: (i) the prolonged exposure to putatively pathogenic isolates (Lazcka et al 2007), (ii) the amount of time and resources needed for each sample (Straub et al 2005), and (iii) the difficulty in obtaining a definitive answer by biochemical tests (Delgado et al 2013).…”

Section: Discussionmentioning

confidence: 99%

“…This rigorous detection scheme is not amenable to the rapid and high-volume screening of multiple environmental samples. Because of these shortcomings, a large body of research has been dedicated to finding rapid and high-throughput screening techniques for the presence of disease-causing organisms in different samples (Girones et al 2010;Noble and Weisberg 2005;Straub et al 2005). One promising and rapidly evolving avenue is biosensors capable of interacting with biological molecules at the nanometer scale (Vikesland and Wigginton 2010).…”

Section: Introductionmentioning

confidence: 99%

“…For the past 20 years the most popular alternative to culture-based detection of pathogens has been molecular-based assays, such as quantitative polymerase chain reaction (qPCR), due to their speed, reliability, and sensitivity (Ishii et al 2013;Straub et al 2005;Yeung et al 2006). However, even molecular-based assays are unreliable when it comes to detection of microorganisms in environmental samples such as wastewater, soil, vegetable surfaces, or aerosols.…”

Section: Introductionmentioning

confidence: 99%

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Rapid MPN-Qpcr Screening for Pathogens in Air, Soil, Water, and Agricultural Produce

Orlofsky

Benami

Gross

et al. 2015

Water Air Soil Pollut

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A sensitive, high-throughput, and costeffective method for screening bacterial pathogens in the environment was developed. A variety of environmental samples, including aerosols, soil of various types (sand, sand/clay mix, and clay), wastewater, and vegetable surface (modeled by tomato), were concomitantly spiked with Salmonella enterica and/or Pseudomonas aeruginosa to determine recovery rates and limits of detection. The various matrices were first enriched with a general pre-enrichment broth in a dilution series and then enumerated by most probable number (MPN) estimation using quantitative PCR for rapid screening of amplicon presence. Soil and aerosols were then tested in non-spiked environmental samples, as these matrices are prone to large experimental variation. Limit of detection in the various soil types was 1-3 colony-forming units (CFU) g −1 ; on vegetable surface, 5 CFU per tomato; in treated wastewater, 5 CFU L −1 ; and in aerosols, >300 CFU mL −1 . Our method accurately identified S. enterica in non-spiked environmental soil samples within a day, while traditional methods took 4 to 5 days and required sorting through biochemically and morphologically similar species. Likewise, our method successfully identified P. aeruginosa in non-spiked aerosols generated by a domestic wastewater treatment system. The obtained results suggest that the developed method presents a broad approach for the rapid, efficient, and reliable detection of relatively low densities of pathogenic organisms in challenging environmental samples.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Rapid MPN-Qpcr Screening for Pathogens in Air, Soil, Water, and Agricultural Produce

Orlofsky

Benami

Gross

et al. 2015

Water Air Soil Pollut

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“…Detection sensitivities of 1 × 10 3 CFU/ml were reported [38]. A multiplex PCR for E. coli O157:H7 and Salmonella with a suspension microarray detection system had a similar sensitivity [40].…”

Section: Microarraysmentioning

confidence: 90%

Recent advances in the development of nucleic acid diagnostics

O’Connor

Glynn

2010

Expert Review of Medical Devices

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“…Improved detection of pathogenic E. coli (Ogunjimi and Choudary, 1999) by immuno-capture PCR, and the sensitive detection of Salmonella (Hoorfar et al, 2000) and Campylobacter (Nogva et al, 2000) by real-time PCR have also been developed; but these procedures are all monospecific and are either laborious or very expensive for routine use in water testing laboratories. More recent improvements have allowed simultaneous detection of several microorganisms in a single assay (Maynard et al, 2005;Straub et al, 2005;Marcelino et al, 2006). The use of such multiplex polymerase chain reaction (m-PCR) has provided rapid and highly sensitive methods for the specific detection of pathogenic microbes in the aquatic environment (Kong et al, 2002).…”

Section: Methods Used In Detection Of Bacterial Pathogens In Watermentioning

confidence: 99%