Comparative advantages and disadvantages of the split G4 DNAzyme-based assay with DNA microarrays, real time reverse-transcription PCR (qRT-PCR), reverse transcription loop-mediated isothermal amplification (RT-LAMP) and semi-quantitative nested PCR.

DENV serotyping assay	Split G4 DNAzyme-based assay50	DNA microarray51	qRT-PCR52,53	RT-LAMP54	Semi-quantitative nested PCR55
Advantages	Inexpensive	Can detect dual infection of two different DENV serotypes	Able to do quantitative measurements	Able to do quantitative measurements	Involves negation of improper primer binding; hence, non-specific detection could be reduced
	Easy to perform and portable		Lower contamination rates due to closed tube operation	Able to do naked-eye visualization
	Amplification-free, lowering risk of target strand contamination		Involves software-driven operation and hence can be applied in high-throughput analysis
	Enables naked-eye visualization
Disadvantages	Unable to perform measurements of target concentrations	Requires specialized and expensive instruments	Requires trained personnel	Complicated design of primers (requires six primers)	Contamination of amplicon products may occur because targets are detected using two sets of primers for a double process of amplification
		Restricted to laboratories with good financial support	Expensive detection equipment and consumables	HPLC purification is needed for two long primers
		Requires trained personnel	Uneconomical for an average laboratory
			Requires fluorescent probes
Requirement of secondary method (agarose gel electrophoresis analysis/ethidium bromide or SYBR green integration)	No	No	No	Optional	Yes
Requirement of thermal cycler/sophisticated instruments	No	Yes	Yes	No	Yes