Rapid and supersensitive allele detection of Plasmodium falciparum chloroquine resistance via a Pyrococcus furiosus argonaute-triggered dual-signal biosensing platform

Table 2 Molecular methods used for identifying mutations in antimalarial drug resistance genes

Classification	Method	LOD	Thermocycler	Advantages	Disadvantages	References
PCR-based assays	mPCR	20 fg/μL	Yes	High accuracy, high sensitivity, time savings, and economic advantages	The quality of the probe is significantly influenced by the base sequence near the detection site	[1]
	HRM	0.3 ng/μl	Yes	Simple, robust, low-cost	Amplicons with sizes greater than 300 bps may produce more errors	[2,3,4]
	AS-PCR	50 parasites/μL	Yes	Most common; easiest	Nonspecific amplification often occurs	[5, 6]
	RFLP	15 parasites/μl	Yes	Rapid, simple, convenient, and inexpensive	Not suitable for large-scale testing	[7,8,9]
Isothermal tests	RPA	3.17 ng/μl	No	No thermocycler required rapid	Susceptible to laboratory contamination; sensitivity not as good as qPCR	[10, 11]
	LFA	3.38 × 10⁵ copies/μL	No	Easy to interpret test results	Susceptible to laboratory contamination	[5, 6, 11]
	RPA-PfAgo	3%	No	No thermocycler needed; rapid; simple to understand the results of the test	The RPA amplification products need to be purified by the purification kit to remove the proteins involved in the RPA reaction
Sequencing methods	Sanger sequencing	20%	Yes	Provide thorough haplotype information	Time-consuming and extensive data analysis requires costly equipment and well-trained technicians	[12]
Sequencing methods	Illumina MiSeq	5%	Yes	Provide thorough haplotype information; faster than Sanger sequencing	Requires expensive equipment and well-trained technicians; and extensive data analysis	[13]

ISSN: 1756-3305