Rapid, label-free genetic detection of enteropathogens in stool without genetic isolation or amplification

Song Han, Mehmet C. Soylu, Ceyhun E. Kirimli, Wei Wu, Bhaswati Sen, Suresh G. Joshi, Christopher Emery, Giang Au, Xiaomin Niu, Richard Hamilton, Kyle Krevolin, Wei Heng Shih, Wan Y. Shih

Research output: Contribution to journalArticle

Abstract

Current genetic detection methods require gene isolation, gene amplification and detection with a fluorescent-tagged probe. They typically require sophisticated equipment and expensive fluorescent probes, rendering them not widely available for rapid acute infection diagnoses at the point of care to ensure timely treatment of the diseases. Here we report a rapid genetic detection method that can detect the bacterial gene directly from patient stools using a piezoelectric plate sensor (PEPS) in conjunction with a continuous flow system with two temperature zones. With stools spiked with sodium dodecyl sulfate (SDS) in situ bacteria lysing and DNA denaturation occurred in the high-temperature zone whereas in situ specific detection of the denatured DNA by the PEPS occurred in the lower-temperature zone. The outcome was a rapid genetic detection method that directly detected bacterial genes from stool in < 40 min without the need of gene isolation, gene amplification, or expensive fluorescent tag but with polymerase chain reaction (PCR) sensitivity. In 40 blinded patient stools, it detected the toxin B gene of Clostridium difficile with 95% sensitivity and 95% specificity. The all-electrical, label-free nature of the detection further supports its potential as a low-cost genetic test that can be used at the point of care.

Original languageEnglish (US)
Pages (from-to)73-80
Number of pages8
JournalBiosensors and Bioelectronics
Volume130
DOIs
StatePublished - Apr 1 2019
Externally publishedYes

Fingerprint

Point-of-Care Systems
Amplification
Labels
Bacterial Genes
Gene Amplification
Genes
Fluorescent Dyes
Temperature
Nucleic Acid Denaturation
DNA
Sodium Dodecyl Sulfate
Clostridium
Denaturation
Polymerase chain reaction
Bacteria
Sensors
Sodium dodecyl sulfate
Costs and Cost Analysis
Sensitivity and Specificity
Equipment and Supplies

Keywords

  • Amplification-free
  • In situ
  • Isolation-free
  • Label-free
  • Piezoelectric plate sensor
  • Stool bacteria genetic detection

ASJC Scopus subject areas

  • Biotechnology
  • Biophysics
  • Biomedical Engineering
  • Electrochemistry

Cite this

Rapid, label-free genetic detection of enteropathogens in stool without genetic isolation or amplification. / Han, Song; Soylu, Mehmet C.; Kirimli, Ceyhun E.; Wu, Wei; Sen, Bhaswati; Joshi, Suresh G.; Emery, Christopher; Au, Giang; Niu, Xiaomin; Hamilton, Richard; Krevolin, Kyle; Shih, Wei Heng; Shih, Wan Y.

In: Biosensors and Bioelectronics, Vol. 130, 01.04.2019, p. 73-80.

Research output: Contribution to journalArticle

Han, S, Soylu, MC, Kirimli, CE, Wu, W, Sen, B, Joshi, SG, Emery, C, Au, G, Niu, X, Hamilton, R, Krevolin, K, Shih, WH & Shih, WY 2019, 'Rapid, label-free genetic detection of enteropathogens in stool without genetic isolation or amplification', Biosensors and Bioelectronics, vol. 130, pp. 73-80. https://doi.org/10.1016/j.bios.2019.01.025
Han, Song ; Soylu, Mehmet C. ; Kirimli, Ceyhun E. ; Wu, Wei ; Sen, Bhaswati ; Joshi, Suresh G. ; Emery, Christopher ; Au, Giang ; Niu, Xiaomin ; Hamilton, Richard ; Krevolin, Kyle ; Shih, Wei Heng ; Shih, Wan Y. / Rapid, label-free genetic detection of enteropathogens in stool without genetic isolation or amplification. In: Biosensors and Bioelectronics. 2019 ; Vol. 130. pp. 73-80.
@article{7dee11d5498f4abbaa72c9acf03ccf06,
title = "Rapid, label-free genetic detection of enteropathogens in stool without genetic isolation or amplification",
abstract = "Current genetic detection methods require gene isolation, gene amplification and detection with a fluorescent-tagged probe. They typically require sophisticated equipment and expensive fluorescent probes, rendering them not widely available for rapid acute infection diagnoses at the point of care to ensure timely treatment of the diseases. Here we report a rapid genetic detection method that can detect the bacterial gene directly from patient stools using a piezoelectric plate sensor (PEPS) in conjunction with a continuous flow system with two temperature zones. With stools spiked with sodium dodecyl sulfate (SDS) in situ bacteria lysing and DNA denaturation occurred in the high-temperature zone whereas in situ specific detection of the denatured DNA by the PEPS occurred in the lower-temperature zone. The outcome was a rapid genetic detection method that directly detected bacterial genes from stool in < 40 min without the need of gene isolation, gene amplification, or expensive fluorescent tag but with polymerase chain reaction (PCR) sensitivity. In 40 blinded patient stools, it detected the toxin B gene of Clostridium difficile with 95{\%} sensitivity and 95{\%} specificity. The all-electrical, label-free nature of the detection further supports its potential as a low-cost genetic test that can be used at the point of care.",
keywords = "Amplification-free, In situ, Isolation-free, Label-free, Piezoelectric plate sensor, Stool bacteria genetic detection",
author = "Song Han and Soylu, {Mehmet C.} and Kirimli, {Ceyhun E.} and Wei Wu and Bhaswati Sen and Joshi, {Suresh G.} and Christopher Emery and Giang Au and Xiaomin Niu and Richard Hamilton and Kyle Krevolin and Shih, {Wei Heng} and Shih, {Wan Y.}",
year = "2019",
month = "4",
day = "1",
doi = "10.1016/j.bios.2019.01.025",
language = "English (US)",
volume = "130",
pages = "73--80",
journal = "Biosensors and Bioelectronics",
issn = "0956-5663",
publisher = "Elsevier Ltd",

}

TY - JOUR

T1 - Rapid, label-free genetic detection of enteropathogens in stool without genetic isolation or amplification

AU - Han, Song

AU - Soylu, Mehmet C.

AU - Kirimli, Ceyhun E.

AU - Wu, Wei

AU - Sen, Bhaswati

AU - Joshi, Suresh G.

AU - Emery, Christopher

AU - Au, Giang

AU - Niu, Xiaomin

AU - Hamilton, Richard

AU - Krevolin, Kyle

AU - Shih, Wei Heng

AU - Shih, Wan Y.

PY - 2019/4/1

Y1 - 2019/4/1

N2 - Current genetic detection methods require gene isolation, gene amplification and detection with a fluorescent-tagged probe. They typically require sophisticated equipment and expensive fluorescent probes, rendering them not widely available for rapid acute infection diagnoses at the point of care to ensure timely treatment of the diseases. Here we report a rapid genetic detection method that can detect the bacterial gene directly from patient stools using a piezoelectric plate sensor (PEPS) in conjunction with a continuous flow system with two temperature zones. With stools spiked with sodium dodecyl sulfate (SDS) in situ bacteria lysing and DNA denaturation occurred in the high-temperature zone whereas in situ specific detection of the denatured DNA by the PEPS occurred in the lower-temperature zone. The outcome was a rapid genetic detection method that directly detected bacterial genes from stool in < 40 min without the need of gene isolation, gene amplification, or expensive fluorescent tag but with polymerase chain reaction (PCR) sensitivity. In 40 blinded patient stools, it detected the toxin B gene of Clostridium difficile with 95% sensitivity and 95% specificity. The all-electrical, label-free nature of the detection further supports its potential as a low-cost genetic test that can be used at the point of care.

AB - Current genetic detection methods require gene isolation, gene amplification and detection with a fluorescent-tagged probe. They typically require sophisticated equipment and expensive fluorescent probes, rendering them not widely available for rapid acute infection diagnoses at the point of care to ensure timely treatment of the diseases. Here we report a rapid genetic detection method that can detect the bacterial gene directly from patient stools using a piezoelectric plate sensor (PEPS) in conjunction with a continuous flow system with two temperature zones. With stools spiked with sodium dodecyl sulfate (SDS) in situ bacteria lysing and DNA denaturation occurred in the high-temperature zone whereas in situ specific detection of the denatured DNA by the PEPS occurred in the lower-temperature zone. The outcome was a rapid genetic detection method that directly detected bacterial genes from stool in < 40 min without the need of gene isolation, gene amplification, or expensive fluorescent tag but with polymerase chain reaction (PCR) sensitivity. In 40 blinded patient stools, it detected the toxin B gene of Clostridium difficile with 95% sensitivity and 95% specificity. The all-electrical, label-free nature of the detection further supports its potential as a low-cost genetic test that can be used at the point of care.

KW - Amplification-free

KW - In situ

KW - Isolation-free

KW - Label-free

KW - Piezoelectric plate sensor

KW - Stool bacteria genetic detection

UR - http://www.scopus.com/inward/record.url?scp=85060934540&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85060934540&partnerID=8YFLogxK

U2 - 10.1016/j.bios.2019.01.025

DO - 10.1016/j.bios.2019.01.025

M3 - Article

VL - 130

SP - 73

EP - 80

JO - Biosensors and Bioelectronics

JF - Biosensors and Bioelectronics

SN - 0956-5663

ER -