Isolation and chemical immobilization of E. coli‐specific bacteriophage with NH2‐MIL‐101(Fe) MOF, a high photoluminescence rod‐shaped microcrystals for low‐level bacteria detection

Abstract

As concern raised by the World Health Organization (WHO) of antibiotic‐resistant and bio‐defensive bacteria, a metal–organic framework (MOF) based optical biosensor came into consideration for precise, quick, and sensitive detection of Escherichia coli (E. coli) (ATCC 10799) using bacteriophage as a bio‐recognition element. In the present study, amine‐functionalized Fe‐based MOF, i.e., NH2‐MIL‐101(Fe), was synthesized by the solvothermal method (approx. 531–1106 nm in size and 20 mV zeta potentials by DLS) and further characterized by SEM, XRD, ATR‐FTIR, UV–VIS, and photoluminescent (PL) spectroscopy. The lytic bacteriophage was isolated from a sewage sample, purified, and concentrated using the ultra‐centrifugation method and achieved a high titer of 7.3 × 1012 PFU/ml. The concentration, stability, and accessible receptor binding domains (RBDs) of the biorecognition element for binding with their analytes play an important role in developing sensitive and specific biosensor systems. To fulfill the mentioned criteria, optimized glutaraldehyde concentration was estimated at 0.25%, at 30 °C for conjugating maximum bacteriophage titer of 8.6 × 105 PFU/ml for each 1 mg amine functionalized iron‐based MOF. The synthesized detection probe has shown excellent photoluminescence and antibacterial activity and achieved a detection limit of 652 CFU/ml over a bacterial detection concentration range from 5.78 × 101 to 5.78 × 106 CFU/ml for E. coli with 10–12 min of response time, high specificity, and long‐term stability even at room temperature. Therefore, it can be inferred that this MOF‐based strategy can be helpful in the specific and sensitive detection of various bacterial pathogens using bacteriophage as a bio‐recognition element.