Table 2.

Advantages and drawbacks of isothermal amplification techniques.

	Advantages	Drawbacks
Rt-qPCR	Specific Sensitive Simultaneous detection and quantification	High cost Specific machine required Absolute quantification relies on standard curve Temperature of reaction
LAMP	Highly specific Can be detected by a cheap turbidimeter No initial heating step Speed (within 1 h)	Complex primers design High temperature (60–65 °C) Possible primer-primer annealing Detection limit
RCA	Simple mechanism Easy primer design Temperature of reaction (30–37 °C) Easily detectable	Numerous rounds of amplifications RNA amplification is complex Works only with circular templates Occurs only linearly over time
NASBA	Specifically designed to detect RNA and in turn RNA viruses Temperature of reaction (41 °C) Fast	Denaturation step Less efficient in amplifying RNA targets out of the range 120–250 bp RNA integrity Pre-heating step
MCDA	Fastest method No initial heating step Speed (40 min) Sensitivity, specificity Low cost	Complex primers design High temperature (60–67 °C)
RPA	High sensitivity Temperature of reaction (37 °C) Simple primers design Extremely quick (20 min) No initial heating step	Non-specific background Possible primer-primer annealing Inhibition by high genomic DNA concentrations Detection limit