Recent advances in molecular diagnostics of hepatitis B virus

Table 1 Comparison between different nucleic acid amplification technologies

Method	Template requirement	Sensitivity/specificity	Enzyme(s) required	Temperature requirement	Primers/primer design	Multiplexpossibility	Rapid detection possibility	Time to detect in (min)	Sensitive to biological inhibitors
PCR	dsDNA1, RNA	About 1-10 copies/Very high	DNA polymerase	95 °C →55-60 °C →68-72 °C (Cyclic)	≥ 2/simple	Yes	Yes2	About 40-120	Yes
LCR	DNA1	About 1-10 copies/Very high	DNA ligase and DNA polymerase	94 °C →65 °C(Cyclic)	4/simple	Yes	Yes2	About 100-180	Yes
LAMP	ssDNA1, RNA	About 5 copies/Very high	Bst DNA polymerase	60-65 °C(Isothermal)	4-6/complex	N/A	Yes234	About 60-90	Less
NASBA	RNA1, DNA	About 1 copy/Very high	Reverse transcriptase, T7 RNA polymerase, RNAse H	37-41 °C(Isothermal)	2/simple	Yes	Yes24	About 60-120	Yes
RCA	Circular ssDNA1	About 10 copies/high	29 DNA polymerase	30-65 °C(Isothermal)	1/simple	N/A	Yes2	About 60-90	Less
TMA	RNA1, DNA	About 1-10 copies/high	Reverse transcriptase, RNA polymerase	50-60 °C(Isothermal)	2/simple	Yes	Yes2	About 120-140	Yes
SDA	ssDNA1, RNA,	About 10 copies/low	Bst DNA polymerase or exo-Klenow Fragment	95 °C →37 °C(Isothermal)	4/complex	Yes	Yes24	About 90-120	Yes
tHDA	dsDNA1	About 1-10 copies/Very high	Helicase, DNA polymerase	60-65 °C(Isothermal)	2/simple	Yes	Yes24	About 75-90	Less

¹Preferred template;

²Real time detection of fluorescence;

³Real time detection of turbidity;

⁴Nucleic acid lateral flow. PCR: Polymerase chain reaction; LCR: Ligase chain reaction; LAMP: Loop mediated isothermal amplification; NASBA: Nucleic acid sequence based amplification; RCA: Rolling circle amplification; TMA: Transcription mediated amplification; SDA: Strand displacement amplification; tHDA: Thermophilic helicase dependent amplification.