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Table 1 Comparative analysis of various isothermal amplification methods

From: Microfluidic chip and isothermal amplification technologies for the detection of pathogenic nucleic acid

Amplification methods

LAMP

NASBA

RCA

HDA

RPA/RAA

Template

DNA and RNA

RNA

DNA and RNA

DNA and RNA

DNA and RNA

Reaction temperature

65°C

37-42°C

37°C

65°C

37-42°C

Amplification time

60 min

90 min

30-240 min

60-120 min

10-30 min

Enzymes for amplification

Bst DNA polymerase

Reverse transcriptase, RNase H, and T7 RNA polymerase

DNA ligase and DNA polymerase

Helicase, single-stranded DNA-binding protein, and DNA polymerase

Recombinase, Bst DNA polymerase, and single-stranded DNA-binding protein

Number of primers

4-6

2

1

2

2

Detection methods for the amplified products

Double-stranded chimeric dye, turbidimetry, indicator lateral flow chromatography, gel electrophoresis, and ELISA

Molecular beacon probes, gel electrophoresis, and ELISA

Fluorescent probes and gel electrophoresis

Gel electrophoresis and fluorescent probes

Fluorescent probes, lateral flow chromatography, and gel electrophoresis

Advantages

Simple reaction system, multiple detection methods, and product detection with naked eyes

Direct detection of RNAs and prevention of contamination

High specificity and low risk of contamination

Low and constant temperature and high sensitivity

Rapid detection, low and constant temperature

Disadvantages

False positivity, complex primer design, and being prone to nonspecific amplification

A necessary preheating step and failure in achieving a real constant temperature

More operating procedures and longer reaction duration

Longer reaction duration

Longer primer probes and being easy to form dimers