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9.3.3 Short description

Detection of biological threats, which could be used against citizens and agricultural produce, involves the recognition of bacteria, viruses and toxins. The identification methods based on nucleic acid and immuno-based methods are used in detecting has been mentioned in the literature. These have also been used in food testing, clinical and environmental applications [50].

 

Table BW.2 - A selection of detection of commonly known bacterial pathogens as cited in the journal paper [50]

 

 

Biorecognition molecules

Target

Type of assay

Estimated time  of assay (h)

Sensitivity (no. of targets)

Specificity

Nucleic acid

pst gene probes and primers

Yersinia pestis

5% nuclease fluorogenic assay

2.5 h

3-30 cells

Very high

 

Heat stable toxin gene (ST-gene) 

E. coli

PCR-ELISA; color amplified PCR (CAPS) 

3.5 h

270 cfu

High

 

Genomic DNA; Escherichia  coli

E. coli and lambda phage ,

Lambda phage Molecular combing, scanning force microscopy

2 h

103 cells

High

 

Universal and specific 16s rRNA primers

B. subtilis, Y. pestis and E. coli 

Multiplex PCR followed by gel electrophoresis

3 h

ND (5 ml of soil)

High

 

 

 

 

 

 

 

 

 

 

Immuno-assays

Fluorescently labeled MAb

Salmonella typhimurium

 Flow cytometry

40 min

103 cells:ml of milk & egg

High

 

Fluorescently labeled PAb to 0157 antigen

E. coli 0157:H7

Antibody-direct Epifluorescent filter technique
(Ab-DEFT)

3 h

104-105 cfu:g feces

High

 

 

Table BW.3 - A selection of recognition techniques for detection of pathogenic viruses as cited in a journal paper [50]

 

 

Biorecognition molecules

Target

Type of assay

Estimated time  of assay (h)

Sensitivity (no. of targets)

Specificity

Nucleic acid

Biotinylated primers for gag gene and internal standard control

HIV-1 RNA:DNA

QC-PCR followed by amplicon capture, probe hybridization and

Luminometry

4h

100

Very high quantitative

 

Biotinylated primers for pol gene

HIV-1 RNA

PCR followed by amplicon capture and colorimetry

3 h

20 virions

Very high

 

Primers for immediate early genes

Active human cytomegalovirus (hCMV)

In situ PCR followed by in situ colorimetry

4 h

100

High

 

Primers for immediate early genes

hCMV

Nested PCR followed by gel electrophoresis

4 h

5 virions

High

 

 

Primers and biotinylated probes

Hepatitis C virus (HCV)

PCR based, digoxigenin labled amplicons are captured by biotinylated probes

3 h

10-100 virions

Very high

 

Digioxigenin labeled riboprobe targeting non-coding sequence

Enteroviruses

Dot blot hybridization and chemiluminescent detection

2.5 h

104-105 TCID

High

 

Digioxigenin labeled Vp1 probe

Polioviruses serotyopes

Dot blot hybridization and chemiluminescent detection

2.5 h

103-104 TCID

High

 

 

 

 

 

 

 

Immuno-assays

Fluorescein labeled human recombinant Fab (rFab)

Cells infected with herpes simplex virus (HSV)

Indirect immunofluorescence

2.5 h

ND

High

 

Fluorescein labeled murine monoclonal Ab to HSV

Cells infected with HSV

Direct immunofluorescence

2 h indirect

ND (less than

High

 

Latex bound antibody to

Cytomegalovirus (CMV)

Latex agglutination, visual assay

10 min

1012

High

 

IgM antibody to

CMV

Enzyme immuno assay

10 min

106

High

 

Antibody

HAV

Immuno electron microscopy

8 h

104-105 virions/ml

Moderate

 

Radio labeled antibody

HAV

Radioimmunoassay

2 h

105-106 virions/ml

Moderate

Self assembled bilayers of Cu2+/L-cysteine have been coated on gold surfaces are used to detect biological agents in microcantilever sensors. Dimethyl methyl phosphonate was used as a sarin nerve gas stimulant in these tests [51].

Label free biosensors also known as optical biosensors are based on direct measurements of a change taking place during a biochemical reaction on the surface of a transducer [52]. Ricin has been detected using optical biosensors within 30 minutes to a detection limit of 10ng/ml. The advantages optical biosensors offer is rapid screening and multi-analyte detection. The main disadvantage for these sensors is the reduced sensitivity for the rapid screening assays [53].

Portable fibre optic biosensors have been demonstrated in the detection of Staphylococcal Enterotoxin B. These biosensors have been demonstrated in the compact, light weight portable
identification system. The multi-channel identification system is based on simultaneous
fluorescence immunoassays on the surface of polystyrene fibre optic probes [54].

NASA has demonstrated the nanotechnology based biosensors that can be used to detect pathogens such as anthrax. These sensors are based on carbon nanofibres and the licensed technology is being commercialised by Early Warning Inc. Electrical signals are measured in
these sensors to identify the presence of a pathogen. These sensors are equipped with
microfludics as well that provides the advantage of field testing, allowing detection within 30 minutes [55].

Aptamers are functional RNA oligonucleotides that are used for sensing biologicial agents. These are also DNA based. Aptamers based biosensors have been reported to be good for security applications. Aptamers based biosensors are relatively immature, in relation to immunoassays, due to the limited availability of aptamers and knowledge of surface immobilization [56]. Single
walled carbon nanotubes based field effect transistors, has been demonstrated to monitor
aptamers - protein affinity binding processes. These offer an advantage over immunological
assays due to their small size in monitoring protein- protein interactions [57].

Array Biosensors have been used to detect targets such as staphylococcal enterotoxin B (SEB), ricin toxin, cholera toxin, mouse IgG and Bacillus globigii (anthrax spore simulant). Prototype for monitoring postal sorting machines was demonstrated to be successful. The array biosensors provide the advantage of being specific and non-destructive [58].

Inhibition of enzyme acetylcholinesterase has been the basis of detecting organophosphate
compounds due to high specificity and selectivity. The development of single thiocholine enzyme based biosensor has been reported in the literature using screen printed carbon electrodes doped with cobalt phthalocyanine. The sensors were observed to be fabricated using electropolymerisation and ablation with ultrasound. Detection limits in the order of 1 * 10-17M were experimentally reported for dichlorvos, parathion and azinphos. In a separate experiment the
same detection limit was reported for paraoxon. The biosensors were reported to have application in environmental monitoring [59,60].

Biosensors based on acetylcholinesterase functionalised carbon nanotubes have been demonstrated in detecting organophosphorus compounds. These sensors have shown excellent limits of detection (0.145 ppb), good precision, electrode to electrode stability, and reproducibility
[61].

Metallic nanowires striped with gold, silver and nickel nanoparticles in a suspended format have been used to identify biological warfare agents such as anthrax, smallpox, ricin and botulinum. Each nanowire relating to a particular antibody detects a pathogen. The made advantage offered by this method of detection is the ability to have up to 100 different striped nanowires which
reduce analysis time significant for multiple analytes [62].

Kane et al. have demonstrated the use of peptide bound carbon nanotubes in detecting toxins such as anthrax, and deactivating the anthrax toxin by using invisible and near infrared light. Carbon nanotubes coatings maybe applied as a thin coating on a range on surfaces [63]. SERS enhancements have been used to detect microorganisms using colloidal metal suspension [64].

Terahertz imaging has been demonstrated for anthrax stimulant bacillus cereus in postal
envelopes [65]. The need for research in detection methods of biological threats was driven by the anthrax spores distribution through the postal services in the United States. Laser induced breakdown spectroscopy (LIBS) is a technique that uses light induced from a laser induced microplasma to determine the composition of the sample, based on elemental and molecular
emission intensity. The main advantages of this technique are high sensitivity, selectivity and
minimal sample preparation. LIBS have been demonstrated to have been effectively used in the detection of bacillus subtilis and ovalbumin. Experimental research has established the
effectiveness of this method with few false positive or false negatives [66].

 

 

 

 

 


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Visits: 1031, Published on: May, 18th 2009, 02:01 PM, Last edit: 2009-05-25 18:27:30 Size: 10 KByte

Tags: biosensors, terahertz, Carbon Nanotubes, nanowires, optical biosensors, nucleic acid based sensing, immunoassay based sensing

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