Fast detection of bacterial gut pathogens on miniaturized devices: an overview

Figures & data

Figure 1. Point-of-care detection methods. Traditional methods for detection of gut pathogens include bacterial cultivation, PCR, biochemistry tests and microscopy analysis and generally take from 24 to 72 hours from sample collection to diagnostic. Point-of-care detection uses immunological tests, molecular biology methods as well as biosensors (colorimetric, electrochemical, fluorometric) leading to a fast diagnostic (generally ranging from 10 minutes to 3 hours). Created using Biorender.com.

Table 1. Examples of immunological methods used for detecting foodborne pathogens.

Pathogen	Detection technique	Sample	LOD	Assay duration	Ref.
Salmonella enteritidis	Nanobody based-ELISA	Spiked milk samples	1.4×10^5 CFU/mL 6 CFU/mL in pre-enriched samples	10 h pre-enrichment 3.5 h ELISA detection	[8]
Salmonella spp.	Label-free Immunochromatographic strips	Poultry samples	10^2 CFU/mL of S.pullorum 10^3 CFU/mL of S.gallinarum 10^4 CFU/mL of S.enterica		[9]
Escherichia coli O157:H7	Paper-based ELISA	Spiked beef sample	10^4 CFU/mL	<3 h	[10]
	Ag enhanced LFIA	Spiked orange juice, milk, & tap water	2 × 10^3 CFU/mL	30 min	[11]
	Antibody coated microspheres	Bacterial suspension	49.1 CFU/μL	1.5 h	[12]
Vibrio parahaemolyticus	magnetic nanobead-based immunoseparation and quantum dot immunofluorescence	Spiked clams	100 CFU/mL	4 h	[13]
	Immunochromatographic strips labelled with nanomaterials	Spiked seafood	5.5 × 105 CFU/mL		[14]

Table 2. Examples of molecular methods used for detecting foodborne pathogens.

Target pathogen	Detection method	Type of sample	LOD	Assay duration	Ref.
Listeria monocytogenes, Escherichia coli, Staphylococcus aureus, Salmonella enterica	mPCR	spiked lettuce and minced meat	10^3 CFU/mL	120 min - mPCR 80 min - agarose gel	[17]
Vibrio parahaemolyticus, Listeria monocytogenes ATCC 19,115, Cronobacter sakazakii, Shigella flexneri, Pseudomonas putida, Escherichia coli O157:H7, Vibrio vulnificus, Vibrio alginolyticus		spiked aquatic food	10^4 CFU/mL	90 min-PCR	[31]
Enterococcu durans KCTC 13,289, Enterococcu faecalis KACC 11,859, Enterococcus faecium KACC 11,954, Enterococcu hirae KACC 16,328, Enterococcu lactis KACC 15,681, Enterococcus mundtii KACC 13,824	qPCR	spiked food samples	10^2 CFU/mL	80 min - PCR	[18]
Campylobacter jejuni, Campilobacter coli	LAMP	broiler production	50 CFU/mL	30 min for amplification	[32]
E. coli O157:H7	Bioluminescent - based LAMP	various food samples	5 CFU/mL	90–120 min	[33]
Listeria monocytogenes	Colorimetric lamp	Contaminated ready-to-eat-food	55 CFU/mL	30 min	[34]
Staphylococcus aureus, Pseudomonas aeruginosa	RPA-LFD	artificially spiked fresh milk	1 CFU/reaction S. aureus and 5 CFU/reaction P.aeruginosa	25 min	[35]
Yersinia enterocolitica	CRISPR/Cas12a-RPA-based system	artificially spiked raw pork meat	1.7 CFU/mL	45 min	[36]

Figure 2. Representation of the various detection methods used in the process of gut pathogen identification and the most common infectious bacteria detected.Created using Biorender.com.

Table 3. Detection of various gut bacteria with multiple types of biosensors.

Type	Biosensor Structure	Target analyte	LOD	Ref.
Resonance	Tungsten oxide-based sensor functionalized with vancomycin	S. aureus	80–100 CFU/mL	[59]
Resonance	AuNPs and MoS2-based LSPR sensor	Shigella	1.56 CFU/mL	[60]
Fluorescence	Quantum dot-based sensor	E. coli, L. monocytogenes, S.Typhimurium	102, 103, and 103 CFU/mL	[61]
Colorimetric	M13 phage-based sensor	E. Coli, P. aeruginosa, V. cholera, X. campestris	100 cells	[62]
Colorimetric	CRISPR-Cas12a and AuNPs based colorimetric sensor	Salmonella	1 CFU/mL	[63]
Colorimetric	Peptide aptamer-based MZI sensor	E. coli	11 CFU/mL	[64]
Piezoelectric	Antibody-AuNPs	Bifidobacterium bifidum	2.1 × 10^2 CFU/mL	[65]
Electrochemical	Molecularly imprinted polymer	TMAO(trimethylamine N-oxide) - metabolite	1–15 ppm	[66]
Electrochemical (Impedance)	OCMCS-Fe3O4 NPs	Campylobacter jejuni	103–107 CFU/mL	[67]
Electrochemical	Bismuth-immobilized carbon nanotube	Helicobacter pylori DNA	0.72–7.92 μg/mL	[68]

Figure 3. Integrated sample-to-answer POCT devices – a. Technical criteria for designing an integrated POCT device; b. Trends and challenges in POCT development and clinical implementation. Created using Biorender.com.

Table 4. Smartphone-based POC tests used for bacterial detection.

POC device	Assay type	Detected bacteria	LOD	Assay duration	Ref.
Smartphone-based fluorescence microscope	Peptide nucleic acid (PNA) based FISH assay	Cronobacter sakazakii, Staphylococcus aureus	≥10^4 CFU/mL	Not specified	[118]
Smartphone-based rt LAMP	Biotin-labeled aptamer	Mycobacterium tuberculosis	10^4 CFU/mL	5 h	[119]
Smartphone-controlled, automated, and portable system	Colorimetric LAMP approach	Staphylococcus aureus	3.2 × 10−3 hemagglutinating units (HAU)/reaction; 30 colony-forming units (CFU)/reaction	40 min	[27]
Smartphone-based detection fluorescence microscopy	Aptamer-functionalized fluorescent magnetic nanoparticles	Staphylococcus aureus	10 CFU/ml	10 min	[120]
Pipette-actuated capillary array comb based on a smartphone-based hand-held device	LAMP	Klebsiella pneumoniae	200 CFU/capillary	85 min	[121]
Smartphone-based rt LAMP	LAMP	urine bacterial analysis	5 × 10^6 CFU/reaction	1 hour	[122]

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