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Premise | Diagnostic method | Sample types | Sensitivity (%) | Specificity (%) | Advantages | Disadvantages | References |
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Detection of virus | Virus isolation (in vivo or in vitro) | Serum, plasma, whole blood, and fresh or FFPE tissues | Variable | 100 | Highly specific | Technical, laborious Requires biosafety level 3 containment May take 1-2 weeks | [1] |
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Detection of viral antigen | ELISA or immunochromatographic assay (ICA) | Serum and CSF | 85 (serum) 80 (CSF) | 89 (serum) 87 (CSF) | Early diagnosis | Commercial assays not widely available Requires biosafety level 3 containment | [16, 17] |
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Detection of viral nucleic acid | RT-PCR | Serum and dried blood spots | 100 | Up to 100 | Highly sensitive and specific Rapid turnaround time Multiplex available | Expensive reagents and specialized equipment |
[13, 16, 18–20] |
Real-time RT-PCR | 100 | Up to 100 | Multiplex available | Expensive reagents and specialized equipment |
Isothermal amplification methods (RT-LAMP) | 100 | 95.25 | Does not require specialized equipment (i.e., thermocyclers) | |
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Detection of host antibody response | ELISA | Serum CSF | IgM: 17 (serum); 48 (CSF) IgG: 45 (serum); 63 (CSF) | IgM: 95 (serum) IgG: 53 (serum) | Widely available Relatively cheaper and easier to perform Rapid bedside tests are available | Possible cross-reactivity with other alphaviruses Elevated IgM does not distinguish recent past infection from acute infection |
[4, 16, 17, 20–22] |
IFA | Serum | 85–97 | 90–98 | Sensitive and specific Commercially available | Lack the ability to quantify antibodies, are subjective, and require special equipment and training |
PRNT | Serum | | | Very specific for alphaviruses; gold standard for confirmation of serologic test results | Requires the use of live virus (requires Biosafety level 3 containment) |
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