Hepatitis B virus subgenotype A1 predominates in liver disease patients from Kerala, India

Table 1 Demographic, clinical and virological characteristics of hepatitis B surface antigen-positive patients with different disease profiles

	Characteristic	Group I HCC (n = 44)	Group II CR (n = 22)	Group III CH (n = 25)	Total (n = 91)
Demographic and clinical data	Gender (M/F)	33/11	18/4	18/7	69/22
	Age (yr) (mean ± SD)	48.70 ± 10.94a	40.68 ± 11.52	26.28 ± 11.10	40.87 ± 14.66
	ALT, IU/L	73.50 ± 11.65	70.50 ± 41.09	56.32 ± 30.01	67.45 ± 37.74
	HBeAg positive1	7 (24.14)	4 (14.3)	9 (37.5)	20 (26.67)
Virological characteristics	HBV DNA log10 copies/mL1	4.79 ± 1.41a	3.38 ± 1.69	3.27 ± 2.12	4.03 ± 1.83
	Number genotyped by direct sequencing/RFLP	21/19	9/12	19/6	86
	Genotype A	33 (82.5)a	14 (66.6)	15 (60)	62 (72.1)
	Subgenotype A1	30	14	15	59 (95)
	Subgenotype A2	3	-	-	3 (5)
	Genotype C	-	1 (4.8)	-	1 (1.2)
	Genotype D	7 (17.5)	6 (28.6)	10 (40)	23 (26.7)
	Subgenotype D12	-	1	3
	Subgenotype D2	4	2	6
	Subgenotype D3	1	-	-

^aP < 0.05 vs chronic hepatitis.

¹Depletion of serum allowed the viral loads to be determined for only 28, and hepatitis B e antigen for 29 HCC sera;

²Subgenotyping of genotype D was performed by direct sequencing. CH: Chronic hepatitis; HCC: Hepatocellular carcinoma; CR: Cirrhosis; SD: Standard deviation; ALT: Alanine aminotransferase; RFLP: Restriction fragment length polymorphism; HBeAg: Hepatitis B e antigen.

Table 2 Multiple logistic regression analysis of basal core promoter/precore region in different disease groups

		HCC		Non HCC (CR/CH)		OR (95%CI)	P value
		Gen A (n = 20)	Gen D (n = 6)	Gen A (n = 26)	Gen D (n = 11)
Basic core promoter/ pre core region	A1762T/G1764A + G1764A only	16 (80)	5 (83)	14 (54)	3 (27)	20.2 (6.3-65)a	0.008aNSd
	C1766T/T1768A	9 (45)	1 (5)	5 (25)	0	25 (7.3-86)a 14.3 (1.7–119)d	0.03a0.05d
	1773T (genotype A)	19 (95)	-	25	-
	1773C (genotype D)	-	4	-	7	-	NSa
	G1862T	18 (90)	2 (33)	21 (81)	1(9)	30.33 (5.62-192.6)d	NSa 0.0001d
	G1896A	0	4 (67)	0	2 (18)	-	NSa 0.0002c
	A1762T/G1764A + G1862T	13 (65)	1 (2)	13 (50)	0	4.81 (0.6-39.4)b	NSa 0.0004b
Complete S region	Pre-S deletions	5 (33.3)e	0	3 (17.6)	1 (9)	1.89 (0.54-6.60)b	NSbNSd

^aComparison between hepatocellular carcinoma (HCC) and non-HCC, all genotypes;

^bComparison between HCC and non-HCC, restricted to genotype A isolates;

^cComparison between HCC and non-HCC, restricted to genotype D isolates;

^dComparison between genotype A and D isolates;

^ePercentage out of 15 genotype A HCC isolates that were sequenced in the S region. CR: Cirrhosis; CH: Chronic hepatitis.

Table 3 Summary of the pre-S mutations prevalent among the three clinical groups and genotypes

Isolate	Age/sex	Clinicalstatus	Subgenotype	PreS1	PreS2					Functionsaffected
				Start codon	Nucleotide from the EcoRI start			Amino acids from preS2 start codon
					Start codon	Deletion size	Position	Deletion size	Position
CHB42	40/M	CH	A1	ATG	ACG	-	-	-	-	A
HCC12	60/M	HCC	A1	ATG	ACG	-	-	-	-	A
HCC25	50/M	HCC	A1	ATG	ACG	-	-	-	-	A
CHB137	16/F	CH	A1	Deletion	ATG	18	2854-2871	6	1-6	I
CHB170	29/F	CH	D2	ATG	ATG	21	35-55	7	16-22	T, B, P
CHB202	33/M	CH	A1	ATG	ATG	24	28-51	8	13-21	T, B, P
CHB413	35/M	CH	A1	ATG	ATG	6	49-54	2	21-22	B, P
HCC29	50/F	HCC	A1	ATG	ATA	33	22-54	11	11-22	A, T, B, P
HCC30	32/M	HCC	A1	ATG	ATG	24	30-53	8	13-21	T, B, P
HCC37	30/F	HCC	A1	ATG	ATG	33	24-56	11	11-22	T, B, P
HCC39	55/M	HCC	A1	ATG	ATG	33	24-56	11	11-22	T, B, P
HCCT3	38/M	HCC	A1	ATG	ATA	54	1-54	18	4-22	A, T, B, P, M

A: PreS2 initiation codon abolished; M1T, M1I; I: PreS1 initiation start codon abolished; M: Morphogenesis domain ps1:103-119 and ps2: 1-4; T: T cell epitope ps1: 109-119/ps2: 1-13; B: B cell epitope, amino acids 14-26; P: Putative neutralizing anti-preS2 antibody, amino acids 1-26; HCC: Hepatocellular carcinoma; CHB: Chronic hepatitis B.