Capture and concentration of viral and bacterial foodborne pathogens using apolipoprotein H

Abstract

The need for improved pathogen separation and concentration methods to reduce time-to-detection for foodborne pathogens is well recognized. Apolipoprotein H (ApoH) is an acute phase human plasma protein that has been previously shown to interact with viruses, lipopolysaccharides (LPS) and bacterial proteins. The purpose of this study was to determine if ApoH was capable of binding and efficiently capturing two representative human norovirus strains (GI.1 and GII.4), a cultivable surrogate, and four bacterial pathogens (Escherichia coli O157:H7, Listeria monocytogenes, Salmonella enterica serovar Enteritidis, and Staphylococcus aureus). Experiments were carried out using an ApoH-conjugated magnetic bead-based capture followed by pathogen detection using nucleic acid amplification. For all three viruses studied, >10% capture efficiency (<1 Log10 loss in RT-qPCR amplifiable units) was observed. The same capture efficiencies were observed for the bacterial pathogens tested, with the exception of E. coli O157:H7 (approximately 1% capture efficiency, or 2 Log10 loss in CFU equivalents). The efficiency of the capture steps did not vary as a consequence of input target concentration or in the presence of an abundance of background microflora. A complementary plate-based capture assay showed that ApoH bound to a variety of human norovirus virus-like particles. ApoH has the potential to be a broadly reactive ligand for separating and concentrating representative foodborne pathogens, both bacteria and viruses.

Detection of Enteric Viruses on Strawberries and Raspberries Using Capture by Apolipoprotein H

“…In addition, we were able to capture more NoV GII.4 from the strawberries using the ApoH method, which suggests that inhibitors other than pectin affect the quantification step, giving a slight advantage to ApoH. It is also important to consider the potential presence of other microorganisms in the HuNoV stocks, such as Gram-negative bacteria, which could compete for ApoH affinity sites and decrease HuNoV recovery [ 15 ]. Different inhibitors could also come from the diluted fecal matter in the HuNoV stock solution.…”

“…Although not large, the difference is significant. The use of the ApoH CaptoVIR kit was described by only one published study, in which only 1 mL of suspension was processed and with only one virus (NoV GI or GII or Tulane virus) per test [ 15 ]. Our recoveries of NoV GII from a virus mixture were slightly lower.…”

“…Apolipoprotein H (ApoH, also known as beta-2-glycoprotein I) is a plasma protein involved in antiphospholipid syndrome (APS syndrome) [ 12 ]. This protein has a broad-spectrum affinity, including for hepatitis B and C viruses [ 13 , 14 ], HuNoVs, and many Gram-positive and Gram-negative bacteria [ 15 ]. To the best of our knowledge, its affinity for HAV is unknown [ 15 ], and the possibility of using it in magnetic bead form for concentrating and detecting the HAV and HuNoVs on berries in a single method should therefore be investigated.…”

New Techniques for the Generation and Analysis of Tailored Microbial Systems on Surfaces

“…These compounds can be either a specific agent designed to capture one particular type of bacteria or general methods that can capture all similar prokaryotes. Depending on the desired application, nonspecific capture agents can improve the capture of bacteria without requiring biofilm formation, improve the adhesion of bacteria to surfaces to facilitate biofilm formation, or concentrate bacteria from dilute samples . The choice of specificity is highly dependent on the desired application.…”

Kaplanmış Manyetik Kürelere DNA Adsorpsiyonu: Fiziksel ve Kimyasal Adsorpsiyonun Karşılaştırılması

“…In the literature, magnetic bead separation processes have been studied for soil clean up (Funada, 2018), wastewater treatment (Lee et al, 2015;Lei et al, 2016;Fuks et al, 2018;Pal et al,. 2018), concentrating pathogenic bacteria (Almand et al, 2016;Lim et al, 2016), viruses (Satoh et al, 2003Almand et al, 2016;Sakudo et al, 2016), and DNA (Satoh et al, 2008;Bordelon et al, 2013). DNA adsorption to the surface of magnetic beads has been mainly used to extract DNA from the sample matrix with higher concentration and lower impurities (Berensmeier, 2006).…”