Locked nucleic acid real-time polymerase chain reaction method identifying two polymorphisms of hepatitis B virus genotype C2 infections, rt269L and rt269I

Table 1 Primers and locked nucleic acid probes developed for the identification of the hepatitis B virus L/I/L2 variants by multiprobe locked nucleic acid real-time polymerase chain reaction

Primer/probe	Sequence (5’ to 3’)1	Tm (°C)2	Target	CH
Primers (product: 128 bp)
Forward	ATGGGATATGTAATTGGAAGtTGGGG	65-67	HBV P gene
Reverse	CCCACAATTCttTGACATACTTTCCAATCAATAGG	67-69	HBV P gene
LNA Probes
L_CTC	5’ 6-FAM-AAA+C+T+CAAR+CA+ATGT - 3’ IABkFQ	61-64	L (WT)	FAM
I_ATC	5’ HEX-AAA+A+T+CAAR+CAA+T+GT - 3’ IABkFQ	61-64	I	HEX
L2_CTA	5’ CY5-AAA+C+T+AAAR+CAA+T+GT - 3’ IABkFQ	60-63	L2	CY5

¹Locked nucleic acid nucleotides are written +A, +C, +T or +G.

²Primer T_m was calculated by using LC PDS software version 2.0, and probe T_m was calculated by https://www.exiqon.com/ls/pages/exiqontmpredictiontool.aspx.

T_m: Melting temperature; CH: Channel; HBV: Hepatitis B virus; WT: Wild type; LNA: Locked nucleic acid.