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

Table 2 Measurement of melting temperatures of the L/I/L2 variants by multiprobe locked nucleic acid real-time polymerase chain reaction

Genotype of positive control DNA [copies, (4.0E + 00)-(4.0E + 08)]	Target sequence1	Measured Tm (°C) in channel2
		Min3	Max3	mean ± SD (detection)3	Min4	Max4		mean ± SD (detection)4	Min5	Max5	mean ± SD (detection) 5
L (n = 34)	AAACTCAAGCAATGTT	61.6	63.1	62.4 ± 0.4 (34, 100%)	-	-		- (0, 0%)	55.2	56.2	55.8 ± 0.2 (26, 76.4%)
L’ (n = 34)	AAACTCAAACAATGTT	57.8	58.7	58.0 ± 0.2 (34, 100%)	-		-	- (0, 0%)	-	-	- (0, 0%)
I (n = 42)	AAAATCAAGCAATGTT	-	-	- (0, 0%)	59.0		61.6	60.2 ± 0.7 (42, 100%)	-	-	- (0, 0%)
I’ (n = 34)	AAAATCAAACAATGTT	-	-	- (0, 0%)	56.3		57.1	56.6 ± 0.2 (34, 100%)	-	-	- (0, 0%)
L2 (n = 34)	AAACTAAAGCAATGTT	-	-	- (0, 0%)	-		-	- (0, 0%)	64.4	64.8	64.6 ± 0.1 (26, 100%)
L2’ (n = 18)	AAACTAAAACAATGTT	-	-	- (0, 0%)	-	-		- (0, 0%)	60.9	61.4	61.2 ± 0.2 (18, 100%)

¹Bold, target codon; underline, A/G polymorphism.

²Bold, target specific melting temperature.

³FAM.

⁴Hex.

⁵Cy5.

Bold words represent genotype-specific T_ms. T_m: Melting temperature; -: No significant melting temperature; SD: Standard deviation.