High level of hepatitis B virus DNA after HBeAg-to-anti-HBe seroconversion is related to coexistence of mutations in its precore and basal core promoter

Table 1 Primers and probes for detections of G1896A or A1762T/G1764A mutations.

Denomination	Sequences (5’-3’)
PCP	BIO -GAGAC TCTAA GGCTT CTCGA TACAG AGCTG AGG
PCMd	DIG -CTCAC GCTAC ATTGT GTGCC TTGGG TGGCT TCA
PCWd	FITC -GTCCG TAGTC TCGTT GTGCC TTGGG TGGCT TGG
BCP1-M	DIG -GCTGA CGATG CGATG GGGAG GAGAT TAGGT TAATG A-3’
BCP1-W	FITC -CGTCC GTAGT GCCGA GGGAG GAGAT TAGGT TAAAG G-3’
PCA	CCCAG CAGAG AATGG CTTGC CTGAG TGCAG TATG
PCSc	CCCGA ATTCC ACCGT GAACG CCCAT CAG
PCAc	CCCAA GCTTG CAGTA TGGTG AGGTG AGCAA TG

BIO: the abbreviation of biotin; DIG: the abbreviation of digoxigenin; FITC: the abbreviation of fluorescein isothiocyanate. The underlined nucleotides had no relationship with HBV.

Table 2 Relationship between CD-PCR results and serum HBV DNA load in 165 serum samples with detectable HBV DNA in serum after HBeAg-to-anti-HBe seroconversion.

	No. (%) of CD-PCR results
	Total G1896A	Total A1762T/G1764A	G1896A only	A1762T/G1764A only	Coexistence
Low-level group (n = 61)	53 (86.9)b	20 (32.8)	35 (57.4)d	2 ( 3.3)	18 (29.5)
Median level group (n = 58)	45 (77.6)	32 (55.2)e	23 (39.6)	10 (17.3)	22 (37.9)
High-level group (n = 46)	30 (65.2)	31 (67.4)f	6 (13.0)	7 (15.2)	24 (52.2)g

¹χ² = 7.08,

^bP<0.01, ²χ² = 21.81,

^dP<0.01, compared with high-level group. ³χ² = 6.06,

^eP<0.05, compared with low-level group. ⁴χ² = 8.20,

^fP<0.01, compared with median level group. ⁵χ² = 5.65,

^gP<0.05, compared with low-level group.

Table 3 Main clinical data of 147 serum samples with G1896A and/or A1762T/G1764A mutations.

	G1896A only (n = 64)	A1762T/G1764A only (n = 19)	Coexistence (n = 64)
Age (yr)	40.5±12.8	43.2±14.2	40.7±12.8
Sex (no. male/no. female)	55/9	18/1	May-59
Alanine aminotransferase (IU/L)	101.9±158.3	121.8±183.1	116.7±137.3
Total bilirubin (mmol/L)	25.2±39.2	24.8±40.6	32.5±55.2b
Serum albumin (g/L)	38.4±5.3	39.1±5.8	37.7±5.3d
Clinical diagnosis
Chronic hepatitis Mild (n)	23	3	13
Median (n)	20	10	14
Gravies (n)	12	4	22f
Liver cirrhosis	9	2	15g

¹t = –2.85,

^bP<0.01, ²t = 6.37,

^dP<0.01, compared with group of G1896A only. ³χ² = 18.3, fP<0.01, ⁴χ² = 4.03,

^gP<0.05, compared with group of mild type of chronic hepatitis B.