Apparatus for two-step surface-enhanced raman spectroscopy

a surface-enhanced raman and raman spectroscopy technology, applied in the field of raman spectroscopy two-step surface-enhanced raman spectroscopy, can solve the problems of culture growth taking several days, pcr taking several hours, and none of the three methods described above can meet the listed requirements, etc., to achieve the effect of sensitivity, speed, and substantial selectivity

Inactive Publication Date: 2016-01-07
REAL TIME ANALYZERS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

Enables the rapid detection of as few as 2500 B. anthracis spores, 100 CFUs of E. coli, and 10 ng / mL of a drug, with enhanced specificity to differentiate between species, achieving significant multiplicative signal enhancement and facilitating at-site measurements.

Problems solved by technology

Unfortunately, none of the three methods described above can meet the listed requirements: culture growth takes several days, PCR takes several hours, and immunoassays lack sensitivity.
In addition, only immunoassays are sufficiently portable for at-site measurements, such as at a military base or a food-processing center.
However, specificity was limited, and bacilli could not be differentiated from clostridia; and differentiation at the species level (e.g. B. cereus versus B. anthracis) was not even a consideration.
Clearly, the basic concept of using SERS as the detector for immunoassays does not provide sufficient sensitivity or speed for the applications discussed above.

Method used

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  • Apparatus for two-step surface-enhanced raman spectroscopy
  • Apparatus for two-step surface-enhanced raman spectroscopy
  • Apparatus for two-step surface-enhanced raman spectroscopy

Examples

Experimental program
Comparison scheme
Effect test

example one

B. Cereus Detected in a Peptide Functionalized Silver-Doped Sol-Gel Coated Capillary

[0101]The silver-doped SER-active sol-gels employed in the filled glass capillaries utilized in this example were prepared in accordance with the method of Farquharson et al. (U.S. Pat. No. 6,623,977), which capillaries were filled with silver-doped sol-gels in accordance with the techniques described by Farquharson et al. (“Rapid dipicolinic acid extraction from Bacillus spores detected by surface-enhanced Raman spectroscopy”Applied Spectroscopy, 4, 351-354, 2004). In essence, a silver amine complex (Ag(NH3)2+OH−), consisting of a 5:1 v / v solution of 1 N AgNO3 and NH4OH (28 wt % NH3 in H2O), was mixed with an alkoxide consisting of a 2:1 v / v solution of methanol and tetramethyl orthosilicate in a 1:8 v / v silver amine:alkoxide ratio. A 10 microL aliquot of the forgoing mixture was then drawn into a 1-mm diameter glass capillary to fill a 10-mm length. After sol-gel formation, the incorporated silver ...

example two

Measurement of E. Coli Detected on an Antibody Functionalized Gold Colloid Coated Glass Slide

Part 8A

[0114]A gold colloid solution was prepared according to Pal et al. (“Photochemically prepared gold nanoparticles: A substrate for surface enhanced Raman scattering,”Current Sci., 84, 1342-1346, 2003). One-inch diameter glass discs were spin-coated at 2500 rpm by adding 1 mL of the gold colloid solution drop-wise to form a monolayer. Once the coating dried, a layer of multi-functional polyethylene glycol ligands—a linker, containing two thiol groups for anchoring onto the gold and a terminal N-hydroxysuccinimide ester group for coupling to antibodies, was added to the surface. After 4 hours the discs were washed with water to remove any unbound PEG from the gold surface. The E. coli selective antibody was then added to the surface and allowed to functionalize over a period of 2 hours, and another buffer wash was thereafter used to remove any unfunctionalized antibody. Then a solution o...

example three

Measurement of Hospital Pathogens: Methicillin Resistant Staphylococcus Aureas (MRSA), Tubercles Bacillus (TB), and Nosocomial Pneumonia (NP)

[0120]As described hereinabove in connection with FIG. 5, a lab-on-a-chip device is designed with three channels, each containing a SER-active metal functionalized with a different binding agent (peptide, aptamer, antibody, etc.) for selectively binding MRSA, TB or NP. In some instances the SER-active metal is chemically attached to the walls of the channels; in other cases it is contained in the channel, preferably incorporated in a sol-gel. A throat sputum sample, collected from a hospital patient, is treated with reagents, such as N-acetyl-L-cysteine and sodium hydroxide, to liquefy the mucans that potentially contain the target hospital pathogens, as described in U.S. Pat. No. 7,713,914. The sample is filtered to trap the pathogens, while passing the mucans and other potentially interfering biochemicals, after which the pathogens are collec...

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Abstract

A kit of components is used in the detection, identification, analysis, and quantitation, by SERS, of a designated target analyte in a sample, comprising: packaging means normally containing a SER-active device component providing a support structure including a SER-active material, a collection component, a container component containing a liquid reagent comprised of a SER-active material, and a component for introducing the liquid reagent into the SER-active device. At least one of the SER-active materials is functionalized with an agent having the specific capability of binding the designated target analyte, and is accessible for the deposit thereon of liquid analyte samples. The SER-active device is constructed for receiving the liquid reagent and for enabling irradiation and collection of Raman-scattered radiation, by and from a Raman spectrometer, cooperatively generated by SER-active metals of the SER-active materials.

Description

BACKGROUND OF THE INVENTION[0001]The ability to measure trace quantities of biological materials in a variety of matrices is important to numerous fields. For example, the measurement of a few Bacillus anthracis spores in air would be invaluable to homeland security, the measurement Escherichia coli in water or Salmonella enterica in food would be invaluable to public safety, and the ability to measure disease causing bacteria (e.g. methicillin resistant Staphylococcus aureus, MRSA), disease biomarkers or nucleotides in body fluids, such as blood, saliva, or urine, would be invaluable to medical diagnosis. Three methods are widely used to detect such trace quantities of biological material, 1) culture growth of bacteria or viruses that are detected by eye with or without the aid of staining and a microscope, 2) immunoassays in which the binding of an antigen to an antibody is detected or, more recently, 3) polymerase chain reactions in which primers are used to separate target nucle...

Claims

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Application Information

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Patent Type & AuthorityApplications(United States)
IPC IPC(8): G01N21/65G01N33/543
CPCG01N33/54373G01N21/658G01N33/58G01N33/587
InventorFARQUHARSON, STUARTSHENDE, CHETAN
OwnerREAL TIME ANALYZERS