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

Han S., Soylu M. Ç., Kirimli C. E., Wu W., Sen B., Joshi S. G., ...Daha Fazla

BIOSENSORS & BIOELECTRONICS, cilt.130, ss.73-80, 2019 (SCI-Expanded) identifier identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 130
  • Basım Tarihi: 2019
  • Doi Numarası: 10.1016/j.bios.2019.01.025
  • Dergi Adı: BIOSENSORS & BIOELECTRONICS
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Sayfa Sayıları: ss.73-80
  • Anahtar Kelimeler: Piezoelectric plate sensor, In situ, Isolation-free, Amplification-free, Label-free, Stool bacteria genetic detection, SURFACE-PLASMON RESONANCE, DIFFICILE-ASSOCIATED DIARRHEA, QUARTZ-CRYSTAL MICROBALANCE, ELECTRICAL DETECTION, ESCHERICHIA-COLI, DNA-DETECTION, COLLABORATIVE PARTNERSHIP, ANTIMICROBIAL STEWARDSHIP, INFECTION PREVENTIONISTS, HUMAN SERUM
  • Erciyes Üniversitesi Adresli: Evet

Özet

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.