Bacteriophage‐receptor binding proteins for multiplex detection of Staphylococcus and Enterococcus in blood

Abstract

Healthcare-associated infections (HCAIs) affect hundreds of millions of patients, representing a significant burden for public health. They are usually associated to multidrug resistant bacteria, which increases their incidence and severity. Bloodstream infections are among the most frequent and life-threatening HCAIs, with Enterococcus and Staphylococcus among the most common isolated pathogens. The correct and fast identification of the etiological agents is crucial for clinical decisionmaking, allowing to rapidly select the appropriate antimicrobial and to prevent from overuse and misuse of antibiotics and the consequent increase in antimicrobial resistance. Conventional culture methods are still the gold standard to identify these pathogens, however, are time-consuming and may lead to erroneous diagnosis, which compromises an efficient treatment. (Bacterio)phage receptor binding proteins (RBPs) are the structures responsible for the high specificity conferred to phages against bacteria and thus are very attractive biorecognition elements with high potential for specific detection and identification of pathogens. Taking into account all these facts, we have designed and developed a new, fast, accurate, reliable and unskilled diagnostic method based on newly identified phage RBPs and spectrofluorometric techniques that allows the multiplex detection of Enterococcus and Staphylococcus in blood samples in less than 1.5 hr after an enrichment step.

The Analysis of OmpA and Rz/Rz1 of Lytic Bacteriophage from Surabaya, Indonesia

“…Phages can use many RBPs to attach to their target bacteria; therefore, finding one does not rule out the possibility of others [ 62 – 64 ]. Santos et al investigated the use of wall teichoic acid as a Staphylococcus phage RBP [ 65 ]. Yan et al looked into the utilization of the colicin A, targeting the outer membrane protein BtuB in E. coli phage [ 66 ], while others looked into FhuA as an RBP in E. coli phage [ 67 ] and phage T5 [ 52 , 68 ].…”

Novel Biorecognition Elements against Pathogens in the Design of State-of-the-Art Diagnostics

“…Bacteriophages (also termed phages) are a type of viruses that infect and replicate within their target bacteria. Due to their high specificity, conferred by receptor binding proteins (RBPs) on the bacteriophage surfaces with which they target bacteria, they have potential for application in diagnostic tools and treatments against bacterial infection [ 149 , 150 , 151 , 152 ]. For instance, Liana et al [ 153 ] functionalised MNPs with a high density of T4 bacteriophages and subsequently used them to capture E. coli.…”

“…Recovery rates ranged from 77.4% to 96.9%, demonstrating good reliability for the detection of bacteria in complex samples [ 151 ]. In an example of multiplex detection [ 150 ], two phage RBPs, gp 18 and gp 109, with potential specificity to the genera Enterococcus or Staphylococcus , respectively, were identified in silico. The RBPs were fused with fluorescent proteins and used in spectrofluorometric assays with the target bacteria for their multiplex detection by fluorescence.…”

Rapid and multiplex detection of nosocomial pathogens on a phage‐based magnetoresistive lab‐on‐chip platform

“…This platform was combined with phage RBPs as bio‐probes for the multiplex detection of nosocomial pathogens, namely Enterococcus and Staphylococcus (Figure 1). For this, we used two RBPs that we have previously reported as being very specific for these pathogens (Santos et al, 2020). The Enterococcus and Staphylococcus cells were firstly marked with MNPs functionalized with the target RBPs (Figure 1a) and, subsequently, they were recognized by RBPs immobilized at the MR chip surface (Figure 1b).…”