Novel assay of competitively differentiated polymerase chain reaction for screening point mutation of hepatitis B virus

Table 1 Primers and probes for detecting of HBV G1896A variant

Denomination	Sequence (5’→3’)
BIO-PCP	BIO-GAGACTCTAAGGCTTCTCGATACAGAGCTGAGG
PCA	GCAGTATGGTGAGGTGAGCAATGCTCAG
DIG-PCMd	DIG-CTCACGCTACATTGTGTGCCTTGGGTGGCTTCA
DIG-PCMc	DIG-TGTGCCTTGGGTGGCTTCA
FLU-PCWd	FLU-GTCCGTAGTCTCGTTGTGCCTTGGGTGGCTTGG
FLU-PCWc	FLU-TGTGCCTTGGGTGGCTTGG
PCS	CCACCGTGAACGCCCATCAG
PCSc	CCCGAATTCCACCGTGAACGCCCATCAG
PCAc	CCCAAGCTTGCAGTATGGTGAGGTGAGCAATG

BIO: the abbreviation of biotin; DIG: the abbreviation of digoxigenin; FLU: the abbreviation of fluorescein.

Table 2 HBV G1896A variant and its relationship with HBV serum markers

	Cases	CD-PCR(%)		CAS-PCR(%)		AS-PCR(%)
		HBV DNA(+)	G1896A variant	HBV DNA(+)	G1896A variant	HBV DNA(+)	G1896A variant
HBsAg(+)/	60	100.0	6.7	100.0	63.3b	93.3	55.0b
HBeAg(+)
HBsAg(+)/	100	62.0c	38.0	59.0	46.0	48.0	40.0
HBeAg(-)
HBsAg(-)/	40	0	0	0	0	0	0
HBeAg(-)

^bP < 0.01, compared with group of CD-PCR, χ² test,

^cP < 0.05, compared with group of AS-PCR, χ² test.

Table 3 Relationship between G1896A variant and HBV DNA level in serum

	HBeAg(+)(n = 56)		HBeAg(-) (n = 48)
	≤ 106copies /ml (n = 14)	> 106 copies /ml(n = 42)	≤ 106 copies /ml(n = 28)	> 106 copies /ml(n = 20)
AS-PCR	2	31	21	19
Competitive	2	35	23	20
AS-PCR
CD-PCR	2	2	21a	8

^aCompared with HBeAg(-) group with a level higher than 10⁶ copies/ml of HBV DNA in serum, Fisher’s exact possibility test, P = 0.0192.