Integration of hepatitis B virus DNA into chromosomal DNA during acute hepatitis B

Table 1 Oligonucleotide primers and PCR cycling profiles used in the study

Fragment	Primer	Position1	Sequence	Denaturation	Annealing	Extension	Size2
A	455(+)	455-474	5’-CAAGGTATGTTGCCCGTTTG-3				1 345
	1 800(–)	1 800-1 773	5’-AGACCAATTTATGCCTACAGCCTCCTA-3’	94 °C 30 s	62 °C 30 s	72 °C 90 s
B	1 687(+)	1 687-1 708	5’-CGACCGACCTTGAGGCATAC-3’
	685(-)	704-685	5’-CGAACCACTGAACAAATGGC-3’	94 °C 30 s	63 °C 30 s	72 °C 30 S	2 198

(+) sense (–) anti-sense.

¹Denotes the nucleotide position of HBV adw genome (GenBank accession no. V00866) where the EcoRI cleavage site is position 1². Size of the amplicons in base pairs.

Table 2 Summary of cloning and sequencing results

Patient details			PCR fragment1	Number of clones					Chromosome number and length
No.	Sex	Age		Sequenced	With vector	With HBV	WithHBV/ chromosomal	With chromosomal
		(yr2)			DNA	DNA only	DNA	DNA only
355	F	18	B	5	1	3	0	1	7 (117 bp)
8225	M	23	A	3	0	2	0	1	1 (278 bp)
			B	4	0	4	0	0	–
28	M	26	A	4	0	2	0	2	4 (512 bp)
									16 (140 bp)
			B	4	0	4	0	0	–
4038	F	27	A	6	0	5	0	1	1 (97 bp)
			B	4	0	4	0	0	–
52833	F	32	A	12	2	6	0	4	4 (456 bp)
									10 (235 bp)
									10 (330 bp)
									2 (422 bp)
			B	4	0	4	0	0	–
962	M	27	A	8	0	4	1	3	1 (300 bp)
									14 (410 bp)
									17 (410 bp)
			B	5	0	5	0	0	–

¹Fragment A amplified with primers 455 (+) and 1 800 (–); fragment B amplified with primers 1 687 (+) and 685 (–).

² yr, years; bp, base pairs; ³Patient with fulminant hepatitis.