|
Technique | Reference | Principle | Advantages | Drawbacks | Current usage status |
|
(A) Fluorescence based methods |
(1) DFA | [24, 25] | Microscopic detection of RSV with specific antibody conjugated with fluorophore. | Easy procedure | Human error, fading of dyes | Research intent, Hospital based procedure, commercial diagnostic assays |
(2) QDs | [26–31] | Detection of signals from fluorescent nanoparticles upon encounter with RSV either through microscopy or flow cytometry | Photostable, inorganic in nature, resistant to metabolic degradation | Toxicity, insolubility | Research intent
|
(3) Molecular beacon based imaging | [32, 33] | Hairpin DNA functionalized gold nanoparticle with fluorophore hybridization with target mRNA | Live cell imaging with real-time detection | Probable gene silencing, metabolic degradation | Research intent |
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(B) Immunoassays |
(1) ELISA | [34, 35] | Specific binding and colorimetric detection of antigen-antibody complex | Easy protocol, high specificity and sensitivity | Cumbersome, prone to human errors | Hospital based procedure, commercial diagnostic assays |
(2) OIA | [36–38] | Presence of specific antigen-antibody complex formed alters the reflective surfaces properties which is visually detected | Easy, rapid, specificity, cost effective | Needs confirmation by other tests for negative samples | Research intent, not commercialized |
(3) LFIA | [39–42] | Immuno-complexes detected chromatographically | Easy, rapid, handy, cost effective, FDA approved | Nonquantitative, limit of sample volume limits detection | Hospital based procedure, commercial diagnostic assays |
|
(C) Molecular methods |
(1) LAMP | [41, 43–45] | Colorimetric/turbidimetric detection of isothermal amplification of DNA using specific primer | Sensitivity and specificity | Semiquantitative, designing compatible primer set | Research intent, not commercialized |
(2) PCR | [46, 47] | Amplification of viral cDNA and visualization of PCR product | Rapid and sensitive than conventional culture methods | High limits of detection | Research intent, hospital based procedure |
(3) Real-Time PCR | [48–53] | Real-time amplification of target DNA or cDNA | Rapid (3–5 hours), highly sensitive and very low limits of detection | Expensive | Research intent, hospital based procedure, commercial assay |
(4) Multiplex PCR | [54–57] | Use of multiple primer and/or probe sets | Simultaneous detection of multiple pathogenic species or strains | Less sensitive | Research intent, hospital based procedure |
(5) Immuno-PCR | [58, 59] | A combination of immunoassay and real-time PCR | Very low limits of detection, improved limits of detection over individual ELISA, and PCR (4000 and 4 fold. respectively) | Complex experimental design | Research intent, not commercialized |
(6) Microarray | [60–74] | Hybridization of sample biomolecules to immobilized target DNA or protein on a chip | Highly sensitive, large scale identification of multiple pathogens; protein and nucleic acid targets | Cost-ineffective | Research intent, hospital based procedure, commercial assay |
|
(D) Biophysical method |
(1) PCR-ESI-MS | [75, 76] | Mass spectroscopy of PCR-amplicons through electron spray dispersion | Highly sensitive and specific even at strain level and efficient multiple pathogens detection. | Expensive | Research intent, not commercialized |
(2) SERS | [77–84] | Inelastic scattering of monochromatic radiation upon interaction with an analyte with low-frequency vibrational and/or rotational energy | Rapid and nondestructive detection of analytes with high sensitivity | Sample preparation | Research intent, not commercialized |
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