Table 1 Differences between molecular detection methods
From: Advances and insights in the diagnosis of viral infections
Method | Molecular methods | ||||||
|---|---|---|---|---|---|---|---|
Nucleic acid amplification | Antibody-antigen detection | CRISPR-Cas | |||||
PCR, real-time PCR | LAMP | NASBA | Elisa | ||||
Enzyme | DNA polymerase | Reverse transcriptase | Antigen | Cas12 | Cas13 | Cas14 | |
Analyte preparation | Purification | Purification and amplification | |||||
Target analyte | RNA, dsDNA (direct pathogen) | RNA | Antibody | ss DNA ds DNA | RNA | ss DNA ds DNA | |
Direct amount of pathogen | Immune response | Direct amount of pathogen | |||||
Additional components | 2 Primers reverse and forward 16–50 bases of complementary nucleic acids; fluorescent reporter and quencher | 4 Unique primers (two inner and two outer); fluorescent reporter and quencher | 2 set of primers and combination of several modification enzymes; fluorescent reporter and quencher | Conversion of enzyme to colorimetric or photoluminescence substrate signal | For ds DNA targets TTTN site; fluorescent reporter and quencher | Depends on RNA secondary structure; fluorescent reporter and quencher | For ds DNA targets TTTA site; fluorescent reporter and quencher |
Non-specific analyte | No | No | DNA | No | ssDNA cleavage | RNA cleavage | ssDNA cleavage |
Additional equipment | Thermocycling | No | No | No | No | ||
Method duration | 2–3 h | 1 h | 2.5 h | 7–8 h | 2 h | ||
Method accuracy | 99–100 | 90 | 93–98 | 92–98 | 95–98 | ||