Hepatitis B virus preS1 deletion is related to viral replication increase and disease progression

Table 2 Primers and HybProbes developed to identify hepatitis B virus preS1 start codon deletion variants by real-time polymerase chain reaction

	Name	Sequence (5'→3')	Tm (°C)1	Target sequence	Channel
Forward primer	HBV_sF	GGGTCACCATATTCTTGGGAAC	62.5	203 - 224 bp of S gene
Reverse primer	HBV_sR	CGAATGCTCCCRCTCCTAC	60.5 or 63.3	203 - 224 bp of S gene
Anchor probe	WT_A	ACAGAAAGATTCGTCCCCATGCCTTGTCGAGG -FL2	72.1
Sensor probe	WT_S	LC Red6403-TGGAAGACGGACCTCCCATG-PH	63.4	Wild type S gene	640
Anchor probe	18D_A	GATTGGGAACAGAAAGATTCGTCCCCATGCC-FL	70.3
Sensor probe	18D_S	LC Red6104-TCGAGGTTTGCTGTAGCT-PH5	59.9	18 bp deletion	610
Anchor probe	21D_A	CCAGAGGATTGGGAACAGAAAGATTCGTCCCC-FL	70.5
Sensor probe	21D_S	LC Red640-GCCTTGTCGAGTGCTGTAGC -PH	64.7	21 bp deletion	640
Anchor probe	15D_A	ACAGAAAGATTCGTCCCCATGCCTTGTCGAGG -FL	73.2
Sensor probe	15D_S	LC Red6706-TTGGTGCTGTAGCTCTT-PH	57	15 bp deletion	670
Anchor probe	preS1_A	GATCCTTGTTGGGGTTGAAGTCCC-FL	65.8
Sensor probe	preS1_S	LC Red6104-ATCTGGATTGTTTGAGTTGGCT-PH	62.4	PreS1	610

¹Temperature was calculated using LC PDS software (version 2.0);

²FL: Fluorescein;

³LC Red640: Light Cycler dye Red640;

⁴LC Red610: Light Cycler dye Red610;

⁵PH: Phosphate;

⁶LC Red670: Light Cycler dye Red670. Tm: Melting temperature; 15D, 18D, 21D_A: 15, 18, 21 base pair deletion anchor probe; 15D, 18D, 21D_S: 15, 18, 21 base pair deletion sensor probe; S gene: Surface region gene.