Rapid single cell detection of Staphylococcus aureus by aptamer-conjugated gold nanoparticles

Staphylococcus aureus is one of the most important human pathogens, causing more than 500,000 infections in the United States each year. Traditional methods for bacterial culture and identification take several days, wasting precious time for patients who are suffering severe bacterial infections. Numerous nucleic acid-based detection methods have been introduced to address this deficiency; however, the costs and requirement for expensive equipment may limit the widespread use of such technologies. Thus, there is an unmet demand of new platform technology to improve the bacterial detection and identification in clinical practice. In this study, we developed a rapid, ultra-sensitive, low cost, and non-polymerase chain reaction (PCR)-based method for bacterial identification. Using this method, which measures the resonance light-scattering signal of aptamer-conjugated gold nanoparticles, we successfully detected single S. aureus cell within 1.5 hours. This new platform technology may have potential to develop a rapid and sensitive bacterial testing at point-of-care.

[1]  C. Tseng,et al.  Rapid detection and identification of clinically important bacteria by high-resolution melting analysis after broad-range ribosomal RNA real-time PCR. , 2006, Clinical chemistry.

[2]  David L. Smith,et al.  Hospitalizations and Deaths Caused by Methicillin-Resistant Staphylococcus aureus, United States, 1999–2005 , 2007, Emerging infectious diseases.

[3]  C. Mirkin,et al.  Nanoparticle-Based Bio-Bar Codes for the Ultrasensitive Detection of Proteins , 2003, Science.

[4]  Classics in infectious diseases. "On abscesses". Alexander Ogston (1844-1929). , 1984, Reviews of infectious diseases.

[5]  J. Szostak,et al.  In vitro selection of RNA molecules that bind specific ligands , 1990, Nature.

[6]  Y. Lin,et al.  Inhibition of multiple thermostable DNA polymerases by a heterodimeric aptamer. , 1997, Journal of molecular biology.

[7]  L. Gold,et al.  Systematic evolution of ligands by exponential enrichment: RNA ligands to bacteriophage T4 DNA polymerase. , 1990, Science.

[8]  Swartz Mn Hospital-acquired infections: diseases with increasingly limited therapies. , 1994 .

[9]  Yajun Song,et al.  [In vitro selection and affinity function of the aptamers to Bacillus anthracis spores by SELEX]. , 2002, Sheng wu hua xue yu sheng wu wu li xue bao Acta biochimica et biophysica Sinica.

[10]  E. Vermaas,et al.  Selection of single-stranded DNA molecules that bind and inhibit human thrombin , 1992, Nature.

[11]  J. Richie,et al.  Targeted nanoparticle-aptamer bioconjugates for cancer chemotherapy in vivo. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[12]  Douglas F. Moore,et al.  Detection and Identification of Mycobacterium tuberculosis Directly from Sputum Sediments by Ligase Chain Reaction , 1998, Journal of Clinical Microbiology.

[13]  Young Keun Kim,et al.  A sensitive method to detect Escherichia coli based on immunomagnetic separation and real-time PCR amplification of aptamers. , 2009, Biosensors & bioelectronics.

[14]  C. Spargo,et al.  Pathogen analysis and genetic predisposition testing using microelectronic arrays and isothermal amplification. , 2000, Journal of investigative medicine : the official publication of the American Federation for Clinical Research.

[15]  A. Cross,et al.  A hospital-wide outbreak of septicemia due to a few strains of Staphylococcus aureus. , 1983, American journal of clinical pathology.

[16]  Kevin W Plaxco,et al.  High specificity, electrochemical sandwich assays based on single aptamer sequences and suitable for the direct detection of small-molecule targets in blood and other complex matrices. , 2009, Journal of the American Chemical Society.

[17]  J. Phair,et al.  Meningitis due to Staphylococcus aureus. , 1985, The American journal of medicine.

[18]  F. Meier,et al.  Evaluation of a Microarray-Based Assay for Rapid Identification of Gram-Positive Organisms and Resistance Markers in Positive Blood Cultures , 2013, Journal of Clinical Microbiology.

[19]  M. Swartz,et al.  Hospital-acquired infections: diseases with increasingly limited therapies. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[20]  Vance G Fowler,et al.  Daptomycin versus standard therapy for bacteremia and endocarditis caused by Staphylococcus aureus. , 2006, The New England journal of medicine.

[21]  B. Swaminathan,et al.  Rapid detection of food-borne pathogenic bacteria. , 1994, Annual review of microbiology.

[22]  Lee-Ann Jaykus,et al.  Selection, characterization, and application of DNA aptamers for the capture and detection of Salmonella enterica serovars. , 2009, Molecular and cellular probes.

[23]  Chengyou Liu,et al.  Selective determination of cysteine by resonance light scattering technique based on self-assembly of gold nanoparticles. , 2006, Analytical biochemistry.

[24]  Konan Peck,et al.  Identification and characterization of oligonucleotides that inhibit Toll‐like receptor 2‐associated immune responses , 2009, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[25]  A. van Belkum,et al.  Nasal carriage of Staphylococcus aureus: epidemiology, underlying mechanisms, and associated risks , 1997, Clinical microbiology reviews.

[26]  Su-Hua Huang,et al.  Detection of Staphylococcus aureus by a sensitive immuno-PCR assay. , 2004, Clinical chemistry.

[27]  J. Moore,et al.  Importance of Staphylococcus aureus in pneumonia in the 1957 epidemic of influenza A. , 1958, Lancet.

[28]  J. Yguerabide,et al.  Light-scattering submicroscopic particles as highly fluorescent analogs and their use as tracer labels in clinical and biological applications. , 1998, Analytical biochemistry.

[29]  F. Luo,et al.  Aptamer from whole-bacterium SELEX as new therapeutic reagent against virulent Mycobacterium tuberculosis. , 2007, Biochemical and biophysical research communications.

[30]  F. Kapral,et al.  Formation of intraperitoneal abscesses by Staphylococcus aureus , 1980, Infection and immunity.

[31]  A. Conde Staphylococcus aureus infections. , 1998, The New England journal of medicine.